Synthetic Methods and Intermediates for Stereoisomeric Compounds Useful for the Treatment of Gastrointestinal and Central Nervous System Disorders

ABSTRACT

The subject invention provides methods and/or processes for making stereoisomeric compounds of formula (X):  
                 
wherein the variables are as defined herein, and compositions for the safe and effective treatment of various gastrointestinal disorders including, but not limited to, gastroparesis, gastroesophageal reflux and related conditions. The compounds of the subject invention are also useful in treating a variety of conditions involving the central nervous system.

This application claims the benefit of U.S. provisional application No.60/713,149, filed Aug. 31, 2005, and U.S. provisional application No.60/747,762, filed May 19, 2006.

BACKGROUND OF INVENTION

Cisapride is one of a class of compounds known as benzamide derivatives,the parent compound of which is metoclopramide. U.S. Pat. Nos. 4,962,115and 5,057,525 (collectively “Van Daele” and incorporated by reference intheir entireties) disclose N-(3-hydroxy-4-piperidenyl) benzamides ofcisapride. Van Daele discloses that these compounds, thepharmaceutically acceptable acid addition salts thereof and thestereochemically isomeric forms thereof, stimulate the motility of thegastrointestinal system.

As a class, these benzamide derivatives have several prominentpharmacological actions. The prominent pharmacological activities of thebenzamide derivatives are due to their effects on the neuronal systemswhich are modulated by the neurotransmitter serotonin. The role ofserotonin, and thus the pharmacology of the benzamide derivatives, hasbeen broadly implicated in a variety of conditions for many years. Thus,research has focused on locating the production and storage sites ofserotonin as well as the location of serotonin receptors in the humanbody in order to determine the connection between these sites andvarious disease states or conditions.

In this regard, it was discovered that a major site of production andstorage of serotonin is the enterochromaffin cell of thegastrointestinal mucosa. It was also discovered that serotonin has apowerful stimulating action on intestinal motility by stimulatingintestinal smooth muscle, speeding intestinal transit, and decreasingabsorption time, as in diarrhea. This stimulating action is alsoassociated with nausea and vomiting.

Because of their modulation of the serotonin neuronal system in thegastrointestinal tract, many of the benzamide derivatives are effectiveanti-emetic agents and are commonly used to control vomiting duringcancer chemotherapy or radiotherapy, especially when highly emetogeniccompounds such as cisplatin are used. This action is almost certainlythe result of the ability of the compounds to block the actions ofserotonin (5HT) at specific sites of action, called the 5HT₃-receptor,which was classically designated in the scientific literature as theserotonin M-receptor. Chemotherapy and radiation therapy may inducenausea and vomiting by the release of serotonin from damagedenterochromaffin cells in the gastrointestinal tract. Release of theneurotransmitter serotonin stimulates both afferent vagal nerve fibers(thus initiating the vomiting reflex) and serotonin receptors in thechemoreceptor trigger zone of the area postrema region of the brain. Theanatomical site for this action of the benzamide derivatives, andwhether such action is central (CNS), peripheral, or a combinationthereof, remains unresolved (Barnes et al., J. Pharm. Pharmacol. 40:586-588, 1988). Cisapride, like the other benzamide derivatives wouldappear to be an effective anti-emetic agent based on its ability tomodulate the activity of serotonin at the 5HT₃ receptor.

A second prominent action of the benzamide derivatives is in augmentinggastrointestinal smooth muscle activity from the esophagus through theproximal small bowel, thus accelerating esophageal and small intestinaltransit as well as facilitating gastric emptying and increasing loweresophageal sphincter tone (Decktor et al., Eur. J. Pharmacol. 147:313-316, 1988). Although the benzamide derivatives are not cholinergicreceptor agonists per se, the aforementioned smooth muscle effects maybe blocked by muscarinic receptor blocking agents such as atropine orneuronal transmission inhibitors of the tetrodotoxin type which affectsodium channels. Similar blocking activity has been reported for thecontractile effects of serotonin in the small intestine. It is currentlybelieved that the primary smooth muscle effects of the benzamidederivatives are the result of an agonist action upon a new class ofserotonin receptors referred to as 5HT₄ receptors which are located oninterneurons in the myenteric plexus of the gut wall. Activation ofthese receptors subsequently enhances the release of acetylcholine fromparasympathetic nerve terminals located near surrounding smooth musclefibers, and it is the combination of acetylcholine with its receptors onsmooth muscle membranes which is the actual trigger for musclecontraction.

A discussion of various 5HT receptors, including the 5HT₄ receptor canbe found in, for example, U.S. Pat. Nos. 6,331,401 and 6,632,827, whichare incorporated by reference herein in their entirety.

Cisapride has been used primarily to treat gastroesophageal refluxdisease (GERD). This disease is characterized as the backward flow ofthe stomach contents into the esophagus. One of the most importantfactors in the pathogenesis of gastroesophageal reflux disease is areduction in the pressure barrier due to the failure of the loweresophageal sphincter. Failure of the lower esophageal sphincter canarise due to a low basal pressure, sphincter relaxation, or to anon-compensated increase in intragastric pressure. Other factors in thepathogenesis of the disease are delayed gastric emptying, insufficientesophageal clearing due to impaired peristalsis and the corrosive natureof the reflux material which can damage esophageal mucosa. Cisapride isthought to strengthen the anti-reflux barrier and improve esophagealclearance by increasing the lower esophageal sphincter pressure andenhancing peristaltic contractions.

Because of its activity as a prokinetic agent, cisapride would alsoappear to be useful to treat dyspepsia, gastroparesis, constipation,post-operative ileus, and intestinal pseudo-obstruction. Dyspepsia is acondition characterized by an impairment of the power or function ofdigestion that can arise as a symptom of a primary gastrointestinaldysfunction or as a complication due to other disorders such asappendicitis, gallbladder disturbances, or malnutrition. Gastroparesisis a paralysis of the stomach brought about by a motor abnormality inthe stomach or as a complication of diseases such as diabetes,progressive systemic sclerosis, anorexia nervosa or myotonic dystrophy.Constipation is a condition characterized by infrequent or difficultevacuation of feces resulting from conditions such as lack of intestinalmuscle tone or intestinal spasticity. Post-operative ileus is anobstruction in the intestine due to a disruption in muscle tonefollowing surgery. Intestinal pseudo-obstruction is a conditioncharacterized by constipation, colicky pain, and vomiting, but withoutevidence of physical obstruction.

Drug toxicity is an important consideration in the treatment of humansand animals. Toxic side effects (adverse effects) resulting from theadministration of drugs include a variety of conditions which range fromlow grade fever to death. Drug therapy is justified only when thebenefits of the treatment protocol outweigh the potential risksassociated with the treatment. The factors balanced by the practitionerinclude the qualitative and quantitative impact of the drug to be usedas well as the resulting outcome if the drug is not provided to theindividual. Other factors considered include the physical condition ofthe patient, the disease stage and its history of progression, and anyknown adverse effects associated with a drug.

Drug elimination is typically the result of metabolic activity upon thedrug and the subsequent excretion of the drug from the body. Metabolicactivity can take place within the vascular supply and/or withincellular compartments or organs. The liver is a principal site of drugmetabolism. The metabolic process can be categorized into synthetic andnonsynthetic reactions. In nonsynthetic reactions, the drug ischemically altered by oxidation, reduction, hydrolysis, or anycombination of the aforementioned processes. These processes arecollectively referred to as Phase I reactions.

In Phase II reactions, also known as synthetic reactions orconjugations, the parent drug, or intermediate metabolites thereof, arecombined with endogenous substrates to yield an addition or conjugationproduct. Metabolites formed in synthetic reactions are, typically, morepolar and biologically inactive. As a result, these metabolites are moreeasily excreted via the kidneys (in urine) or the liver (in bile).Synthetic reactions include glucuronidation, amino acid conjugation,acetylation, sulfoconjugation, and methylation.

More than 90% of a dose of cisapride is metabolized by oxidativeN-dealkylation at the piperidine nitrogen or by aromatic hydroxylationoccurring on either the 4-fluorophenoxy or benzamide rings.

The administration of cisapride to a human has been found to causeserious adverse effects including CNS disorders, increased systolicpressure, interactions with other drugs, diarrhea, and abdominalcramping. Further, it has been reported that intravenous administrationof cisapride demonstrates the occurrence of additional adverse effectsnot experienced after oral administration of cisapride (Stacher et al.[1987] Digestive Diseases and Sciences 32(11):1223-1230). It is believedthat these adverse effects are caused by the metabolites that resultfrom the oxidative dealkylation or aromatic hydroxylation of thecompound which occurs in the cytochrome P450 detoxification system.Cisapride is also subject to a number of undesirable drug/druginteractions that are also a result of metabolism by the cytochrome P450system.

Between July 1993 and December 1999, cisapride (PROPULSID, JanssenPharmaceutica Products, L.P.) was reportedly associated with at least341 serious cardiac arrhythmias. These arrhythmias include ventriculartachycardia, ventricular fibrillation, torsades de pointes, and QTprolongation. Eighty (80) deaths have been reported. As a result ofthese adverse effects, the product was voluntarily withdrawn from theopen market in the United States; however, the drug is available throughan investigational limited access program.

The safety of 5HT₄ receptor agonists with gastrointestinal (GI)prokinetic activity has been limited due to cardiac effects(prolongation of QTc intervals, tachycardia, torsades de pointes) andadverse drug interactions due to hepatic cytochrome P-450 metabolism. AGI prokinetic agent of this class that lacks these liabilities would bevery valuable in several therapeutic areas including GERD and gastricemptying disorders. Certain cisapride derivatives have been described inU.S. Pat. No. 6,552,046 and WO 01/093849 (incorporated by referenceherein in their entireties), however further compounds with even moreadvantageous properties would be desirable.

It has now been discovered that certain stereoisomers of one suchesterified structural and/or functional analog of cisapride havedistinct and particularly advantageous properties.

BRIEF SUMMARY OF THE INVENTION

The subject invention provides methods and processes for makingcompounds and compositions of formula (X), as well as the intermediatesuseful in preparing the compounds of formula (X), for the safe andeffective treatment of various gastrointestinal disorders including, butnot limited to, gastroparesis, gastroesophageal reflux and relatedconditions. The compounds of the subject invention are also useful intreating a variety of conditions involving the central nervous system.

The compounds of the invention comprise compounds of formula X:

and pharmaceutically acceptable salts thereof, wherein

the bonds at positions 3 and 4 are cis relative to each other;

L is —(C₁-C₆ alkyl)-(in one aspect, —(C₃-C₅ alkyl)—), —(C₁-C₆alkyl)-C(O)-, or -C(O)-(C₁-C₆ alkyl)-, wherein each of the alkyl groupsis optionally substituted with 1 or 2 groups that are independentlyhalogen, C₁-C₄ alkoxy, or OH and wherein one carbon in the alkyl portionof L may be replaced by —N(R₉)—;

R₁ is halogen;

R₂ is amino, NH(C₁-C₄ alkyl) or N(C₁-C₄ alkyl)(C₁-C₄ alkyl);

R₃ is OH or C₁-C₄ alkoxy;

R is H or methyl; and

R₅ is —O—C₃-C₈ cycloalkyl, —O-heterocycloalkyl, heterocycloalkyl, aryl,—O-aryl, —N(R))—(C₀-C₆ alkyl)-C(O)-aryl, or —N(R))—C₀-C₆ alkyl-aryl,—O-heteroaryl, —N(R₉)—C₁-C₆(O)-heteroaryl, or —N(R₉)—C₀-C₆alkyl-heteroaryl, wherein each of the cyclic groups is unsubstituted orsubstituted at one or more substitutable positions with C₁-C₆ alkyl,C₁-C₆ alkoxy, halogen, C₁-C₆ haloalkyl, C₁-C₆ haloalkoxy, hydroxyl,hydroxy-C₁-C₄-alkyl, amino, —NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)(C₁-C₆alkyl), —(C₀-C₆ alkyl)-C(O)R₁₁, or —O—(C₀-C₆ alkyl)-C(O)R₁₁,methylsulfone, C₀-C₆-sulfonamide, or NO₂; wherein

R₉ at each occurrence is independently H or C₁-C₄ alkyl;

R₁₁ is C₁-C₆ alkyl, OH, or

R₁₁ is C₁-C₆ alkoxy, optionally substituted with 1 or 2 groups that areindependently C₁-C₄ alkoxy, amino, —NH(C₁-C₆ alkyl), —N(C₁-C₆alkyl)(C₁-C₆ alkyl), —(C₀-C₆ alkyl)-C(O)N(R₉)-heterocycloalkyl,—O-heterocycloalkyl, —C₁-C₆(O)N(R₉)-heteroaryl, or heteroaryl, wherein

-   -   the heterocycloalkyl groups are optionally substituted with 1,        2, or 3 groups that are independently halogen, C₁-C₆ alkyl,        C₁-C₆ alkoxy, hydroxy, hydroxy C₁-C₆ alkyl, C₁-C₆        alkoxycarbonyl, —CO₂H, CF₃, or OCF₃,    -   the heteroaryl group is optionally substituted with 1, 2, or 3        groups that are independently halogen, C₁-C₆ alkyl, C₁-C₆        alkoxy, hydroxy, hydroxy C₁-C₆ alkyl, C₁-C₆ alkoxycarbonyl,        —CO₂H, CF₃, or OCF₃; or

R₁₁ is —O-heterocycloalkyl wherein the heterocycloalkyl is optionallysubstituted with 1, 2, or 3 groups that are independently halogen, C₁-C₆alkyl, C₁-C₆ alkoxy, hydroxy, hydroxy C₁-C₆ alkyl, C₁-C₆ alkoxycarbonyl,—CO₂H, CF₃, or OCF₃; and

R₂₀ is C₁-C₆ alkoxy (preferably C₁-C₄ alkoxy, more preferably methoxy),or OH.

The invention also encompasses compositions comprising at least onecompound of formula (X) made by the methods and/or processes of theinvention and at least one pharmaceutically acceptable excipient,adjuvant, carrier, or solvent.

The compounds of formula (X) that are made by the methods and/orprocesses of the invention are useful in the treatment or prevention ofgastroesophageal reflux disease and substantially reduce adverse effectsassociated with the administration of cisapride. These adverse effectsinclude, but are not limited to, diarrhea, abdominal cramping andelevations of blood pressure and heart rate.

Additionally, the compounds and compositions made by the methods and/orprocesses of the invention are useful in treating emesis and otherconditions, including but not limited to dyspepsia, gastroparesis,constipation, post-operative ileus and intestinal pseudo-obstruction. Asan added benefit, adverse effects associated with the administration ofcisapride are also reduced in these methods of treatment.

Advantageously, the compounds made by the methods and/or processes ofthe invention are ligands for the 5HT₄ receptor and, accordingly, can beused to treat conditions mediated through this receptor. These receptorsare located in several areas of the central nervous system and themodulation of these receptors can be used to effect desired modulationsof the CNS.

Advantageously, compounds made according to the methods and/or processesof the subject invention for making stereoisomeric compounds generallycontain an ester moiety that does not detract from the ability of thesecompounds to provide a therapeutic benefit, but which makes them moresusceptible to degradation by serum and/or cytosolic esterases, therebyavoiding the cytochrome P450 drug detoxification system associated withadverse effects caused by cisapride and reducing the incidence of suchadverse events.

The subject invention further provides methods of treatment comprisingthe administration of the compounds of formula (X) made utilizing themethods and/or processes of the invention in therapeutically effectiveamounts to individuals in need of treatment for gastroesophageal refluxdisease, dyspepsia, gastroparesis, constipation, post-operative ileus,and intestinal pseudo-obstruction; and related conditions.

Advantageously, the therapeutic compounds made utilizing the methodsand/or processes of the subject invention are stable in storage andprovide for safer metabolism of the drugs as compared to other drugs;therefore, the compounds of the subject invention can be used with alower incidence of side effects and toxicity.

In a further aspect, the subject invention pertains to the breakdownproducts (preferably metabolic breakdown products), which are formedwhen the therapeutic compounds made by the methods and/or processes ofthe subject invention are acted upon by esterases. These breakdownproducts can be used as described herein to monitor the clearance of thetherapeutic compounds from a patient.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph representing the Concentration-Response Curves for5-HT₄ Receptor Agonism of ATI-7505, serotonin, Cisapride, and ATI-7500.

FIG. 2 is a graph representing gastric emptying in fed dogs. The datashown are normalized to the averaged vehicle control times of MMC returnvalues. Values represent mean +SEM of 5 dogs. *p<0.05 versus vehiclecontrols

FIG. 3 is a graph representing the metabolism of ATI-7505 and ATI-7500,with and without the CYP450 dependent Cofactor, NADPH. The plots showmean and SD μM concentrations of ATI-7505 and ATI-7500. ATI-7505 (2 μM)was incubated with human microsomal protein (1 mg) in the presence orabsence of NADPH regenerating system (cofactor).

DETAILED DESCRIPTION OF THE INVENTION

In a further aspect, the invention provides methods and/or processes formaking compounds of Formula (X), as well as intermediates useful inpreparing the compounds of formula (X), wherein

-   -   R₅ is —O—C₃-C₈ cycloalkyl, —O-heterocycloalkyl,        heterocycloalkyl, wherein the heterocycloalkyl group is selected        from piperidinyl, piperazinyl, pyrrolidinyl, aza-bicyclo-octyl,        in certain embodiments aza-bicyclo[2.2.2]octyl,        aza-bicyclo[3.2.1]octyl, aza-bicyclo-nonyl, aza-bicyclo-decyl,        indolinyl, morpholinyl, thiomorpholinyl,        S,S-dioxothiomorpholinyl, and imidazolidinyl, —O-aryl,        —N(R₉)—C(O)-aryl, or —N(R₉)—C₀-C₆ alkyl-aryl, wherein each of        the cyclic groups is unsubstituted or substituted at one or more        substitutable positions with C₁-C₆ alkyl, C₁-C₆ alkoxy, halogen,        C₁-C₆ haloalkyl, C₁-C₆ haloalkoxy, hydroxyl,        hydroxy-C₁-C₄-alkyl, amino, —NH(C₁-C₆ alkyl), —N(C₁-C₆        alkyl)(C₁-C₆ alkyl), —C(O)R₁₁, or NO₂; wherein    -   R₉ at each occurrence is independently H or C₁-C₄ alkyl; and    -   R₁₁ is C₁-C₆ alkyl, OH, or

R₁₁ is C₁-C₆ alkoxy, optionally substituted with 1 or 2 groups that areindependently C₁-C₄ alkoxy, amino, —NH(C₁-C₆ alkyl), —N(C₁-C₆alkyl)(C₁-C₆ alkyl), —C(O)N(R₉)-heterocycloalkyl, heterocycloalkyl orheteroaryl, wherein

-   -   the heterocycloalkyl group is selected from pyrrolidinyl,        piperidinyl, piperazinyl, morpholinyl, aza-bicyclo-octyl, in        certain embodiments aza-bicyclo[2.2.2]octyl,        aza-bicyclo[3.2.1]octyl, aza-bicyclo-nonyl and        aza-bicyclo-decyl, wherein the heterocycloalkyl groups are        optionally substituted with 1, 2, or 3 groups that are        independently halogen, C₁-C₆ alkyl, C₁-C₆ alkoxy, hydroxy,        hydroxy C₁-C₆ alkyl, C₁-C₆ alkoxycarbonyl, —CO₂H, CF₃, or OCF₃,    -   the heteroaryl group is selected from pyridyl, pyrimidyl,        quinolinyl, isoquinolinyl, and indolyl, wherein the heteroaryl        groups are optionally substituted with 1, 2, or 3 groups that        are independently halogen, C₁-C₆ alkyl, C₁-C₆ alkoxy, hydroxy,        hydroxy C₁-C₆ alkyl, C₁-C₆ alkoxycarbonyl, —CO₂H, CF₃, or OCF₃;        or    -   R₁₁ is —O-heterocycloalkyl wherein the heterocycloalkyl is        selected from piperidinyl, pyrrolidinyl, imidazolidinyl,        morpholinyl, aza-bicyclo-octyl, in certain embodiments        aza-bicyclo[2.2.2]octyl, aza-bicyclo[3.2.1]octyl,        aza-bicyclo-nonyl, aza-bicyclo-decyl, and tetrahydrofuranyl, and        wherein each heterocycloalkyl group is optionally substituted        with 1, 2, or 3 groups that are independently halogen, C₁-C₆        alkyl, C₁-C₆ alkoxy, hydroxy, hydroxy C₁-C₆ alkyl, C₁-C₆        alkoxycarbonyl, —CO₂H, CF₃, or OCF₃.

In another aspect, the invention provides methods and/or processes formaking compounds of Formula (X), as well as intermediates useful inpreparing the compounds of formula (X), wherein R₁ is chloro.

In yet another aspect, the invention provides methods and/or processesfor making compounds of Formula (X), as well as intermediates useful inpreparing the compounds of formula (X), wherein R₂ is amino.

In still another aspect, the invention provides methods and/or processesfor making compounds of Formula (X), as well as intermediates useful inpreparing the compounds of formula (X), wherein R₃ is methoxy.

In another aspect, the invention provides methods and/or processes formaking compounds of Formula (X), as well as intermediates useful inpreparing the compounds of formula (X), wherein R₄ is H or methyl.

In still yet another aspect, the invention provides methods and/orprocesses for making compounds of Formula (X), as well as intermediatesuseful in preparing the compounds of formula (X), wherein R₁ is chloro;R₂ is amino; R₃ is methoxy; and R₄ is H or methyl.

In yet another aspect, the invention provides methods and/or processesfor making compounds of Formula (X), as well as intermediates useful inpreparing the compounds of formula (X), wherein R₁ is chloro; R₂ isamino; R₃ is methoxy; R is H, and L is —(C₄-C₆ alkyl)-C(O)—.

In another aspect, the invention provides methods and/or processes formaking compounds of formula (X), as well as intermediates useful inpreparing the compounds of formula (X), wherein two or more previouslydescribed aspects are combined.

In another aspect, the invention provides methods and/or processes formaking compounds of Formula (XI), which are compounds of formula (X)wherein L is —(CH₂)₅—C(O)—:

In yet still another aspect, the invention provides methods and/orprocesses for making compounds of formula (XI), as well as intermediatesuseful in preparing the compounds of formula (XI), wherein R₁ is chloro;R₂ is amino; R₃ is methoxy; and R is H or methyl.

In still another aspect, the invention provides methods and/or processesfor making compounds of formula (XI), as well as intermediates useful inpreparing the compounds of formula (XI), wherein R₅ is—O-heterocycloalkyl, wherein the heterocycloalkyl group is selected fromaza-bicyclo-octyl, in certain embodiments 1-aza-bicyclo[2.2.2]oct-3-ylor 8-aza-bicyclo[3.2.1]oct-3-yl, aza-bicyclo-nonyl, aza-bicyclo-decyl,where the aza nitrogen, is optionally substituted with methyl or ethyl;and R is H or methyl.

In still yet another aspect, the invention provides methods and/orprocesses for making compounds of formula (XI), as well as intermediatesuseful in preparing the compounds of formula (XI), wherein R₅ is—O-heterocycloalkyl, wherein the heterocycloalkyl group is selected frompiperidinyl, piperazinyl, or pyrrolidinyl, each of which isunsubstituted or substituted at one or two positions with groups thatare independently C₁-C₄ alkyl, C₁-C₄ alkoxy, halogen, C₁-C₄ haloalkyl(in one aspect, CF₃), C₁-C₄ haloalkoxy (in one aspect OCF₃), hydroxyl,hydroxy C₁-C₄ alkyl, amino, —NH(C₁-C₄ alkyl), —N(C₁-C₄ alkyl)(C₁-C₄alkyl), —(C₁-C₆ alkyl)-C(O)R₁₁, or NO₂; and R is H or methyl.

In yet another aspect, the invention provides methods and/or processesfor making compounds of formula (XI), as well as intermediates useful inpreparing the compounds of formula (XI), wherein R₅ is—O-heterocycloalkyl, wherein the heterocycloalkyl group is selected fromindolinyl, morpholinyl, thiomorpholinyl, S,S-dioxothiomorpholinyl, andimidazolidinyl, each of which is unsubstituted or substituted at one ortwo positions with groups that are independently C₁-C₄ alkyl, C₁-C₄alkoxy, halogen, C₁-C₄ haloalkyl (in one aspect, CF₃), C₁-C₄ haloalkoxy(in one aspect OCF₃), hydroxyl, hydroxy C₁-C₄ alkyl, amino, —NH(C₁-C₄alkyl), —N(C₁-C₄ alkyl)(C₁-C₄ alkyl), —(C₀-C₆ alkyl)-C(O)R₁₁, or NO₂;and R is H or methyl.

In yet another aspect, the invention provides methods and/or processesfor making compounds of formula (XI), as well as intermediates useful inpreparing the compounds of formula (XI), wherein R₅ is —O-phenyl,N(R₉)—(C₀-C₆ alkyl)-C(O)-phenyl, or —N(R₉)—C₀-C₄ alkyl-phenyl, whereinthe phenyl group is substituted with one or two groups that areindependently C₁-C₄ alkyl, C₁-C₄ alkoxy, halogen, C₁-C₄ haloalkyl (inone aspect, CF₃), C₁-C₄ haloalkoxy (in one aspect OCF₃), hydroxyl,hydroxy C₁-C₄ alkyl, amino, —NH(C₁-C₄ alkyl), —N(C₁-C₄ alkyl)(C₁-C₄alkyl), —(C₀-C₆ alkyl)-C(O)R₁₁, or NO₂; and R₄ and R₉ are independentlyH or methyl.

In another aspect, the invention provides methods and/or processes formaking compounds of formula (XI), as well as intermediates useful inpreparing the compounds of formula (XI), wherein R is H.

In yet another aspect, the invention provides methods and/or processesfor making compounds of formula (XI), as well as intermediates useful inpreparing the compounds of formula (X), wherein R₁₁ is C₁-C₆ alkoxy,optionally substituted with 1 or 2 groups that are independently C₁-C₄alkoxy, amino, —NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)(C₁-C₆ alkyl), —(C₀-C₆alkyl)-C(O)N(R₉)-heterocycloalkyl, or heterocycloalkyl wherein theheterocycloalkyl group is selected from pyrrolidinyl, piperidinyl,piperazinyl, and morpholinyl, wherein the heterocycloalkyl groups areoptionally substituted with 1, 2, or 3 groups that are independentlyhalogen, C₁-C₆ alkyl, C₁-C₆ alkoxy, hydroxy, hydroxy C₁-C₆ alkyl, C₁-C₆alkoxycarbonyl, —CO₂H, CF₃, or OCF₃.

In another aspect, the invention provides methods and/or processes formaking compounds of formula (XI), as well as intermediates useful inpreparing the compounds of formula (XI), wherein two or more previouslydescribed aspects are combined.

In another aspect, the invention provides methods and/or processes formaking compounds of Formula (XII), i.e., compounds of formula (X) of thefollowing formula, as well as intermediates useful in preparing thecompounds of formula (XII):

-   -   wherein R₁₅ is H, C₁-C₆ alkyl, C₁-C₆ alkoxy, halogen, C₁-C₆        haloalkyl (in one aspect CF₃), C₁-C₆ haloalkoxy (in one aspect        OCF₃), hydroxyl, hydroxy C₁-C₄ alkyl, amino, —NH(C₁-C₆ alkyl),        —N(C₁-C₆ alkyl)(C₁-C₆ alkyl), methylsulfone, C₀-C₆-sulfonamide        or NO₂, and R₁₆ is H or —O—(C₀-C₆ alkyl)-C(O)R₁₁. In another        aspect, R₁₅ is H.

In yet another aspect, the invention provides methods and/or processesfor making compounds of formula (XII), as well as intermediates usefulin preparing the compounds of formula (XII), wherein R₄ and R₉ areindependently H or methyl and R₁₁ is OH.

In still yet another aspect, the invention provides methods and/orprocesses for making compounds of formula (XII), as well asintermediates useful in preparing the compounds of formula (XII),wherein R₄ and R₉ are independently H or methyl and R₁₁ is C₁-C₆ alkoxy,optionally substituted with 1 or 2 groups that are independently C₁-C₄alkoxy, amino, —NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)(C₁-C₆ alkyl), —(C₀-C₆alkyl)-C(O)N(R₉)-heterocycloalkyl, or heterocycloalkyl wherein theheterocycloalkyl group is selected from aza-bicyclo-octyl, in certainembodiments 1-aza-bicyclo[2.2.2]oct-3-yl or8-aza-bicyclo[3.2.1]oct-3-yl, aza-bicyclo-nonyl, aza-bicyclo-decyl,where the aza nitrogen is optionally substituted with methyl or ethyl,pyrrolidinyl, piperidinyl, piperazinyl, and morpholinyl, wherein theheterocycloalkyl groups are optionally substituted with 1, 2, or 3groups that are independently halogen, C₁-C₆ alkyl, C₁-C₆ alkoxy,hydroxy, hydroxy C₁-C₆ alkyl, C₁-C₆ alkoxycarbonyl, —CO₂H, CF₃, or OCF₃,and R₄ and R₉ are independently H or methyl. In another aspect, R₄, R₉,and R₁₁ are as previously defined and R₁₅ is H, R₁ is chloro; R₂ isamino; and R₃ is methoxy.

In yet still another aspect, the invention provides methods and/orprocesses for making compounds of formula (XII), as well asintermediates useful in preparing the compounds of formula (XII),wherein R₄ and R₉ are independently H or methyl and R₁₁ is C₁-C₆ alkoxy,optionally substituted with 1 or 2 groups that are independently C₁-C₄alkoxy, amino, —NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)(C₁-C₆ alkyl), orheteroaryl, wherein the heteroaryl group is selected from pyridyl,pyrimidyl, quinolinyl, isoquinolinyl, and indolyl, wherein theheteroaryl groups are optionally substituted with 1, 2, or 3 groups thatare independently halogen, C₁-C₆ alkyl, C₁-C₆ alkoxy, hydroxy, hydroxyC₁-C₆ alkyl, C₁-C₆ alkoxycarbonyl, —CO₂H, CF₃, or OCF₃; and R₄ and R₉are independently H or methyl. In another aspect, R₄, R₉, and R₁₁ are aspreviously defined and R₁₅ is H, R₁ is chloro; R₂ is amino; and R₃ ismethoxy.

In still another aspect, the invention provides methods and/or processesfor making compounds of formula (XII), as well as intermediates usefulin preparing the compounds of formula (XII), wherein at least one of R₄and R₉ is H.

In another aspect, the invention provides methods and/or processes formaking compounds of formula (XII), as well as intermediates useful inpreparing the compounds of formula (XII), wherein two or more previouslydescribed aspects are combined.

In another aspect, the invention provides methods and/or processes formaking compounds of Formula (XIII), i.e., compounds of formula (X) ofthe following formula, as well as intermediates useful in preparing thecompounds of formula (XIII):

-   -   wherein R₁₅ is H, C₁-C₆ alkyl, C₁-C₆ alkoxy, halogen, C₁-C₆        haloalkyl (in one aspect CF₃), C₁-C₆ haloalkoxy (in one aspect        OCF₃), hydroxyl, hydroxy C₁-C₄ alkyl, amino, —NH(C₁-C₆ alkyl),        —N(C₁-C₆ alkyl)(C₁-C₆ alkyl), or methylsulfone,        C₀-C₆-sulfonamide, NO₂, and R₁₆ is H or —O—(C₀-C₆        alkyl)-C(O)R₁₁. In another aspect, R₁₅ is H.

In yet another aspect, the invention provides methods and/or processesfor making compounds of formula (XIII), as well as intermediates usefulin preparing the compounds of formula (XIII), wherein

-   -   R₄ and R₉ are independently H or methyl, and R₁₁ is OH, C₁-C₄        alkoxy (in another aspect, C₁-C₃ alkoxy), or C₁-C₂ alkoxy-C₁-C₃        alkoxy-. In another aspect, R₄, R₉, and R₁₁ are as previously        defined and R₁ is chloro; R₂ is amino; and R₃ is methoxy.

In still yet another aspect, the invention provides methods and/orprocesses for making compounds of formula (XIII), as well asintermediates useful in preparing the compounds of formula (XIII),wherein R₄ and R₉ are independently H or methyl, and R₁₁ is C₁-C₄ alkoxysubstituted with amino, —NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)(C₁-C₆ alkyl),aza-bicyclo-octyl, in certain embodiments 1-aza-bicyclo[2.2.2]oct-3-ylor 8-aza-bicyclo[3.2.1]oct-3-yl, aza-bicyclo-nonyl, aza-bicyclo-decyl,where the aza nitrogen is optionally substituted with methyl or ethyl;and R₄ is H or methyl, pyrrolidinyl, piperidinyl, morpholinyl, pyridyl,or —(C₀-C₆ alkyl)-C(O)NH-pyrid-4-yl. In another aspect, R₄, R₉, and R₁₁are as previously defined and R₁ is chloro; R₂ is amino; and R₃ ismethoxy.

In still another aspect, the invention provides methods and/or processesfor making compounds of formula (XIII), as well as intermediates usefulin preparing the compounds of formula (XIII), wherein R₄ and R₉ areindependently H or methyl, and R₁₁ is C₁-C₄ alkoxy substituted withamino, —NH(C₁-C₆ alkyl), or —N(C₁-C₆ alkyl)(C₁-C₆ alkyl). In anotheraspect, R₄, R₉, and R₁₁ are as previously defined and R₁ is chloro; R₂is amino; and R₃ is methoxy.

In yet another aspect, the invention provides methods and/or processesfor making compounds of formula (XIII), as well as intermediates usefulin preparing the compounds of formula (XIII), wherein R₄ and R₉ areindependently H or methyl, and R₁₁ is C₁-C₄ alkoxy substituted withpyrrolidinyl, piperidinyl, morpholinyl, pyridyl, or —(C₀-C₆alkyl)-C(O)NH-pyrid-4-yl. In another aspect, R₄, R₉, and R₁₁ are aspreviously defined and R₁ is chloro; R₂ is amino; and R₃ is methoxy.

In still another aspect, the invention provides methods and/or processesfor making compounds of formula (XIII), as well as intermediates usefulin preparing the compounds of formula (XIII), wherein at least one of R₄and R₉ is H.

In another aspect, the invention provides methods and/or processes formaking compounds of formula (XIII), as well as intermediates useful inpreparing the compounds of formula (XIII), wherein two or morepreviously described aspects are combined.

In another aspect, the invention provides methods and/or processes formaking compounds of formula (XIV), i.e., compounds of formula (X) of thefollowing formula, as well as intermediates useful in preparing thecompounds of formula (XIV):

-   -   wherein R₁₅ is H, C₁-C₆ alkyl, C₁-C₆ alkoxy, halogen, C₁-C₆        haloalkyl (in one aspect CF₃), C₁-C₆ haloalkoxy (in one aspect        OCF₃), hydroxyl, hydroxy C₁-C₄ alkyl, amino, —NH(C₁-C₆ alkyl),        —N(C₁-C₆ alkyl)(C₁-C₆ alkyl), methylsulfone, C₀-C₆-sulfonamide,        or NO₂, and R₁₆ is H or —O—(C₀-C₆ alkyl)-C(O)R₁₁. In another        aspect, R₁₅ is H.

In still another aspect, the invention provides methods and/or processesfor making compounds of formula (XIV), as well as intermediates usefulin preparing the compounds of formula (XIV), wherein R₄ and R₉ areindependently H or methyl, and R₁₁ is OH, C₁-C₄ alkoxy (in anotheraspect, C₁-C₃ alkoxy) or C₁-C₂ alkoxy-C₁-C₃ alkoxy-. In another aspect,R₄, R₉, and R₁₁ are as previously defined and R₁ is chloro; R₂ is amino;and R₃ is methoxy. In still another aspect, at least one of R₄ and R₉ isH.

In yet still another aspect, the invention provides methods and/orprocesses for making compounds of formula (XIV), as well asintermediates useful in preparing the compounds of formula (XIV),wherein R₄ and R₉ are independently H or methyl, and R₁₁ is C₁-C₄ alkoxysubstituted with amino, —NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)(C₁-C₆ alkyl),aza-bicyclo-octyl, in certain embodiments 1-aza-bicyclo[2.2.2]oct-3-ylor 8-aza-bicyclo[3.2.1]oct-3-yl, aza-bicyclo-nonyl, aza-bicyclo-decyl,where the aza nitrogen is optionally substituted with methyl or ethyl;and R₄ is H or methyl, pyrrolidinyl, piperidinyl, morpholinyl, pyridyl,or —(C₀-C₆ alkyl)-C(O)NH-pyrid-4-yl. In another aspect, R₄, R₉, and R₁₁are as previously defined and R₁ is chloro; R₂ is amino; and R₃ ismethoxy.

In still another aspect, the invention provides methods and/or processesfor making compounds of formula (XIV), as well as intermediates usefulin preparing the compounds of formula (XIV), wherein R₄ and R₉ areindependently H or methyl, and R₁₁ is C₁-C₄ alkoxy substituted withamino, —NH(C₁-C₆ alkyl), or —N(C₁-C₆ alkyl)(C₁-C₆ alkyl). In anotheraspect, R₄, R₉, and R₁₁ are as previously defined and R₁ is chloro; R₂is amino; and R₃ is methoxy.

In yet another aspect, the invention provides methods and/or processesfor making compounds of formula (XIV), as well as intermediates usefulin preparing the compounds of formula (XIV), wherein R₄ and R₉ areindependently H or methyl, and R₁₁ is C₁-C₄ alkoxy substituted withpyrrolidinyl, piperidinyl, morpholinyl, pyridyl, or —(C₀-C₆alkyl)-C(O)NH-pyrid-4-yl. In another aspect, R₄, R₉, and R₁₁ are aspreviously defined and R₁ is chloro; R₂ is amino; and R₃ is methoxy.

In still another aspect, the invention provides methods and/or processesfor making compounds of formula (XIV), as well as intermediates usefulin preparing the compounds of formula (XIV), wherein at least one of R₄and R₉ is H.

In another aspect, the invention provides methods and/or processes formaking compounds of formula (XIV), as well as intermediates useful inpreparing the compounds of formula (XIV), wherein two or more previouslydescribed aspects are combined.

In another aspect, the invention provides methods and/or processes formaking compounds of formula (XV), i.e., compounds of formula (X) of thefollowing formula, as well as intermediates useful in preparing thecompounds of formula (XV):

-   -   wherein n is 1 or 2.

In still another aspect, the invention provides methods and/or processesfor making compounds of formula (XV), as well as intermediates useful inpreparing the compounds of formula (XV), wherein R₄ is H or methyl, andR₁₁ is OH, C₁-C₄ alkoxy (in another aspect, C₁-C₃ alkoxy) or C₁-C₂alkoxy-C₁-C₃ alkoxy-. In another aspect, R₄ and R₁₁ are as previouslydefined and R₁ is chloro; R₂ is amino; and R₃ is methoxy. In stillanother aspect, at least one of R₄ and R₉ is H.

In yet still another aspect, the invention provides methods and/orprocesses for making compounds of formula (XV), as well as intermediatesuseful in preparing the compounds of formula (XV), wherein R₄ and R₉ areindependently H or methyl, and R₁₁ is C₁-C₄ alkoxy substituted withamino, —NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)(C₁-C₆ alkyl),aza-bicyclo-octyl, in certain embodiments 1-aza-bicyclo[2.2.2]oct-3-ylor 8-aza-bicyclo[3.2.1]oct-3-yl, aza-bicyclo-nonyl, aza-bicyclo-decyl,where the aza nitrogen is optionally substituted with methyl or ethyl;and R₄ is H or methyl, pyrrolidinyl, piperidinyl, morpholinyl, pyridyl,or —C(O)NH-pyrid-4-yl. In another aspect, R₄, R₉, and R₁₁ are aspreviously defined and R₁ is chloro; R₂ is amino; and R₃ is methoxy.

In still another aspect, the invention provides methods and/or processesfor making compounds of formula (XV), as well as intermediates useful inpreparing the compounds of formula (XV), wherein R₄ and R₉ areindependently H or methyl, and R₁₁ is C₁-C₄ alkoxy substituted withamino, —NH(C₁-C₆ alkyl), or —N(C₁-C₆ alkyl)(C₁-C₆ alkyl). In anotheraspect, R₄, R₉, and R₁₁ are as previously defined and R₁ is chloro; R₂is amino; and R₃ is methoxy.

In yet another aspect, the invention provides methods and/or processesfor making compounds of formula (XV), as well as intermediates useful inpreparing the compounds of formula (XV), wherein R₄ is H or methyl, andR₁₁ is C₁-C₄ alkoxy substituted with aza-bicyclo-octyl, in certainembodiments 1-aza-bicyclo[2.2.2]oct-3-yl or8-aza-bicyclo[3.2.1]oct-3-yl, aza-bicyclo-nonyl, aza-bicyclo-decyl,where the aza nitrogen is optionally substituted with methyl or ethyl;and R₄ is H or methyl, pyrrolidinyl, piperidinyl, morpholinyl, pyridyl,or —(C₀-C₆ alkyl)-C(O)NH-pyrid-4-yl. In another aspect, R₄, R₉, and R₁₁are as previously defined and R₁ is chloro; R₂ is amino; and R₃ ismethoxy.

In another aspect, the invention provides methods and/or processes formaking compounds of formula (XV), as well as intermediates useful inpreparing the compounds of formula (XV), wherein two or more previouslydescribed aspects are combined.

In another aspect, the invention provides methods and/or processes formaking compounds, as well as intermediates useful in preparing thecompounds, according to any one of formulas (X), (XI), (XII), (XIII),(XIV) or (XV), wherein R₁, R₂, and R₃ are oriented on the phenyl ring asfollows:

In another aspect, the invention provides methods and/or processes formaking compounds, as well as intermediates useful in preparing thecompounds, according to any one of formulas (X), (XI), (XII), (XIII),(XIV) or (XV), wherein bond 3 has the “S” configuration and bond 4 hasthe “R” configuration.

In still another aspect, the invention provides methods and/or processesfor making compounds, as well as intermediates useful in preparing thecompounds, according to any one of formulas (X), (XI), (XII), (XIII),(XIV) or (XV), wherein R₁, R₂, and R₃ are oriented on the phenyl ring asfollows:

-   -   and bond 3 has the “S” configuration and bond 4 has the “R”        configuration.

In another aspect, the invention provides methods and/or processes formaking compounds, as well as intermediates useful in preparing thecompounds, according to any one of formulas (X), (XI), (XII), (XIII),(XIV) or (XV), wherein bond 3 has the “R” configuration and bond 4 hasthe “S” configuration.

In another aspect, the invention provides methods and/or processes formaking compounds, as well as intermediates useful in preparing thecompounds, according to any one of formulas (X), (XI), (XII), (XIII),(XIV) or (XV), wherein R₁, R₂, and R₃ are oriented on the phenyl ring asfollows:

-   -   and bond 3 has the “R” configuration and bond 4 has the “S”        configuration.

In still another aspect, the invention provides methods and/or processesfor making compounds, as well as intermediates useful in preparing thecompounds, of formula (X), wherein R₁ is chloro; R₂ is amino; R₃ ismethoxy; R₄ is H, and R₁, R₂, and R₃ have the following orientation onthe phenyl ring:

and

-   -   L is —(C₃-C₅ alkyl)- wherein one carbon may be replaced by        —N(R₉)—, or —(C₂-C₆ alkyl)-C(O)—. In yet another aspect, the R1,        R2, and R3 are as defined and oriented on the phenyl ring as        previously described, R₄ is as previously defined and R₅ is        —O-heterocycloalkyl, wherein the heterocycloalkyl group is        selected from aza-bicyclo-octyl, in certain embodiments        1-aza-bicyclo[2.2.2]oct-3-yl or 8-aza-bicyclo[3.2.1]oct-3-yl,        aza-bicyclo-nonyl, aza-bicyclo-decyl, where the aza nitrogen is        optionally substituted with methyl or ethyl, piperidinyl,        piperazinyl, and pyrrolidinyl, wherein the piperidinyl,        piperazinyl, and pyrrolidinyl groups are unsubstituted or        substituted at one or two positions with groups that are        independently C₁-C₄ alkyl, C₁-C₄ alkoxy, halogen, C₁-C₄        haloalkyl, C₁-C₄ haloalkoxy, hydroxyl, hydroxy C₁-C₄ alkyl,        amino, —NH(C₁-C₄ alkyl), —N(C₁-C₄ alkyl)(C₁-C₄ alkyl), —(C₀-C₆        alkyl)-C(O)R₁₁, or NO₂, wherein

R₁₁ is C₁-C₆ alkoxy, optionally substituted with 1 or 2 groups that areindependently C₁-C₄ alkoxy, amino, —NH(C₁-C₆ alkyl), —N(C₁-C₆alkyl)(C₁-C₆ alkyl), —(C₀-C₆ alkyl)-C(O)N(R₉)-heterocycloalkyl, orheterocycloalkyl wherein the heterocycloalkyl group is selected fromaza-bicyclo-octyl, in certain embodiments 1-aza-bicyclo[2.2.2]oct-3-ylor 8-aza-bicyclo[3.2.1]oct-3-yl, aza-bicyclo-nonyl, aza-bicyclo-decyl,where the aza nitrogen is optionally substituted with methyl or ethyl;and R₄ is H or methyl, pyrrolidinyl, piperidinyl, piperazinyl, andmorpholinyl, wherein the heterocycloalkyl groups are optionallysubstituted with 1, 2, or 3 groups that are independently halogen, C₁-C₆alkyl, C₁-C₆ alkoxy, hydroxy, hydroxy C₁-C₆ alkyl, C₁-C₆ alkoxycarbonyl,—CO₂H, CF₃, or OCF₃.

In still yet another aspect, the invention provides methods and/orprocesses for making compounds, as well as intermediates useful inpreparing the compounds, of formula (X), wherein

-   -   R₁ is chloro; R₂ is amino; R₃ is methoxy; R₄ is H, and R₁, R₂,        and R₃ have the following orientation on the phenyl ring:        and    -   L is —(C₃-C₅ alkyl)- wherein one carbon may be replaced by        —N(R₉)—, or —(C₂-C₆ alkyl)-C(O)—. In yet another aspect, the R₁,        R₂, and R₃ are as defined and oriented on the phenyl ring as        previously described, R₄ is as previously defined and R₅ is        heterocycloalkyl, which is selected from aza-bicyclo-octyl, in        certain embodiments 1-aza-bicyclo[2.2.2]oct-3-yl or        8-aza-bicyclo[3.2.1]oct-3-yl, aza-bicyclo-nonyl,        aza-bicyclo-decyl, where the aza nitrogen, is optionally        substituted with methyl or ethyl.

In still yet another aspect, the invention provides methods and/orprocesses for making compounds of formula (X), as well as intermediatesuseful in preparing the compounds of formula (X), wherein

-   -   R₁ is chloro; R₂ is amino; R₃ is methoxy; R₄ is H, and R₁, R₂,        and R₃ have the following orientation on the phenyl ring:        and    -   L is —(C₃-C₅ alkyl)- wherein one carbon may be replaced by        —N(R₉)—, or —(C₂-C₆ alkyl)-C(O)—. In yet another aspect, the R₁,        R₂, and R₃ are as defined and oriented on the phenyl ring as        previously described, R₄ is as previously defined and R₅ is        —N(R₉)—C₀-C₄ alkyl-aryl or —N(R₉)-(CO-C₆ alkyl)-C(O)-aryl,        wherein the aryl group is unsubstituted or substituted at one or        more substitutable positions with C₁-C₆ alkyl, C₁-C₆ alkoxy,        halogen, C₁-C₆ haloalkyl, C₁-C₆ haloalkoxy, hydroxyl,        hydroxyalkyl, amino, —NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)(C₁-C₆        alkyl), —(C₀-C₆ alkyl)-C(O)R₁₁, or NO₂. In still another aspect,        the aryl group is a phenyl substituted with —(C₀-C₆        alkyl)-C(O)R₁₁ and optionally substituted with 1 or 2 groups        independently selected from C₁-C₆ alkyl, C₁-C₆ alkoxy, halogen,        CF₃, OCF₃, hydroxyl, hydroxyalkyl, amino, —NH(C₁-C₄ alkyl),        —N(C₁-C₄ alkyl)(C₁-C₄ alkyl), or NO₂, wherein    -   R₁₁ is C₁-C₆ alkoxy, optionally substituted with 1 or 2 groups        that are independently C₁-C₄ alkoxy, amino, —NH(C₁-C₆ alkyl),        —N(C₁-C₆ alkyl)(C₁-C₆ alkyl), —(C₀-C₆        alkyl)-C(O)N(R₉)-heterocycloalkyl, or heterocycloalkyl wherein        the heterocycloalkyl group is selected from pyrrolidinyl,        piperidinyl, piperazinyl, and morpholinyl, wherein the        heterocycloalkyl groups are optionally substituted with 1, 2, or        3 groups that are independently halogen, C₁-C₆ alkyl, C₁-C₆        alkoxy, hydroxy, hydroxy C₁-C₆ alkyl, C₁-C₆ alkoxycarbonyl,        —CO₂H, CF₃, or OCF₃. In a preferred aspect the —(C₀-C₆        alkyl)-C(O)R₁₁ group is attached to position 4 of the phenyl        ring.

In still another aspect, the orientation of bonds 3 and 4 of a compoundmade according to the methods and/or processes of the invention is asfollows:

In a preferred aspect, the orientation of bonds 3 and 4 of a compoundmade according to the methods or processes of the invention is asfollows:

The invention further provides methods for treating emesis, dyspepsia,gastroparesis, constipation, intestinal pseudo-obstruction,gastroesophageal reflux, or post-operative ileus, the method comprisingadministering a therapeutically effective amount of a compound or saltaccording of formula (X) that is made according to the methods and/orprocesses of the invention to a patient in need of such treatment.

The subject invention provides methods and/or processes for makingcompounds that are more susceptible to degradation by serum and/orcytosolic esterases than cisapride, thus avoiding the adverse effectsassociated with metabolism by cytochrome P450.

Advantageously, the therapeutic compounds made according to the methodsand/or processes of the subject invention are stable in storage but havea relatively short half-life in the physiological environment;therefore, the compounds of the subject invention can be used with alower incidence of side effects and toxicity.

In a preferred aspect of the subject invention, therapeuticstereoisomeric compounds that are made according to the methods and/orprocesses of the invention are provided, which are useful in thetreatment of gastroesophageal reflux disease and that contain an estergroup, which is susceptible to degradation by esterases, therebybreaking down the compound and facilitating its efficient removal fromthe treated individual. In a preferred aspect, the therapeuticstereoisomeric compounds are metabolized by the Phase I drugdetoxification system.

A further aspect of the subject invention pertains to the breakdownproducts (preferably metabolic breakdown products, i.e., metabolites,generally acids of parent esters) that are produced when the therapeuticcompounds made by the methods and/or processes of the subject inventionare acted upon by an esterase. The presence of these breakdown productsin the urine or serum can be used to monitor the rate of clearance ofthe therapeutic compound from a patient.

Degradation of the compounds made according to the methods and/orprocesses of the subject invention by esterases is particularlyadvantageous for drug metabolism because these enzymes are ubiquitouslydistributed and their activity is not dependent on age, gender, ordisease state to the same extent as oxidative hepatic drug metabolism.

The subject invention further provides methods of treating disorders,such as gastroesophageal reflux disease comprising the administration ofa therapeutically effective amount of at least one stereoisomericstructural and/or functional analog of cisapride to an individual inneed of treatment. In a specific aspect, the subject invention providesstereoisomeric structural and/or functional analogs of cisapride andpharmaceutical compositions of these esterified compounds.

The subject invention further provides materials and methods for thetreatment of emesis and such other conditions, including but not limitedto dyspepsia, gastroparesis, constipation, and intestinalpseudo-obstruction, while substantially reducing adverse effectsassociated with the administration of cisapride.

In a preferred aspect of the subject invention, methods and/or processesfor making therapeutic stereoisomeric compounds are provided, whichcompounds are useful in the treatment of gastroesophageal reflux,dyspepsia, gastroparesis, constipation, post-operative ileus, andintestinal pseudo-obstruction and which contain an ester group which isacted upon by esterases thereby breaking down the compound andfacilitating its efficient removal from the treated individual.

The subject invention further provides methods of synthesizing theadvantageous compounds of the subject invention. Particularly, methodsof producing and purifying such stereoisomeric compounds are taught.Methods of adding such ester moieties and of producing and purifyingstereoisomers, are well known to the skilled artisan and can be readilycarried out utilizing the guidance provided herein.

Preferred Compounds

In a preferred aspect, the present invention provides methods and/orprocesses for making isolated stereoisomers of Compound I, as well asintermediates useful in preparing the stereoisomers, which containsthree chiral centers.

Two of the chiral centers exist in cisapride and norcisapride and are inthe cis configuration in the active drugs:

Thus, for example, pharmaceutically active norcisapride is a racemicmixture of the two cis enantiomers:

In one aspect, the current invention is particularly concerned withproviding methods and/or processes for making compounds with aparticular configuration at the third chiral center, in thequinuclidinol moiety, as well as intermediates useful in preparing suchcompounds. This group is eliminated in the conversion to the acidmetabolite referred to herein as ± Compound II:

While Compound I stereoisomers can be made by conjugating R or Squinuclidinol to (+)- or (−)-norcisapride, giving Compounds III, IV, Vand VI, preferred methods are described below and do not utilize acisapride core.

In a preferred aspect, the subject invention pertains to methods and/orprocesses for making stereoisomerically isolated compounds, as well tothe intermediates useful in preparing such compounds, and compositionscomprising such compounds. The isolated stereoisomeric forms of thecompounds made according to the methods and/or processes of theinvention are substantially free from one another (i.e., instereoisomeric excess). In other words, the “R” forms of the compoundsare substantially free from the “S” forms of the compounds and are,thus, in stereoisomeric excess of the “S” forms. Conversely, “S” formsof the compounds are substantially free of “R” forms of the compoundsand are, thus, in stereoisomeric excess of the “R” forms. In one aspectof the invention, the isolated stereoisomeric compounds made accordingto the methods and/or processes of the invention are in at least about80% stereoisomeric excess. In a preferred aspect, the compounds madeaccording to the methods and/or processes of the invention are in atleast about 90% stereoisomeric excess. In a more preferred aspect, thecompounds made according to the methods and/or processes of theinvention are in at least about 95% stereoisomeric excess. In an evenmore preferred aspect, the compounds made according to the methodsand/or processes of the invention are in at least about 97.5%stereoisomeric excess. In a most preferred aspect, the compounds madeaccording to the methods and/or processes of the invention are in atleast about 99% stereoisomeric excess. Similarly, the “(+)” and “(−)”forms of the compounds are also provided in stereoisomeric excess.

As described herein, the various stereoisomers made according to themethods and/or processes of the invention have particular unexpectedproperties that, advantageously, can be used to customize treatment fora particular set of circumstances. Thus, for example, compoundscontaining the (3′R)-isomer in the quinuclidinyl ester moiety, i.e.,compounds III and IV, are rapidly metabolized by esterases in humanplasma, whereas compounds containing the (3′S)-isomer of quinuclidinol,i.e., compounds V and VI, undergo a much slower metabolism.

Thus, the (3′R)-isomers of compound I can be used when a short-durationof action is preferred, for example stimulation of gastric motility inan acute episode, such as pulsatile administration to patients withacute gastroparesis, or in acute gastroesophageal reflux. Anotheradvantage of rapid metabolism by esterases to an substantially lessactive metabolites, i.e., compound II, is the very low probability ofdrug-drug interactions and toxicity. Therefore these short-acting(R)-isomers can be advantageously used as an intravenous formulation fortreating gastroesophageal reflux in premature newborns, which arenotoriously unable to metabolize drugs as well as adults because theirCYP450 system is not fully developed. In these newborn, a drug havingrapid metabolism by a system other than CYP450, e.g., esterases, is agreat advantage. On the other hand, the (3′S)-isomers of compound I madeaccording to the methods and/or processes of the invention are best usedin chronic situations of the same ailments, for example gastroparesis indiabetic patients or cancer patients under opiates, or in chronicgastroesophageal reflux in patients who need 24-hour coverage.

In addition to their differences in metabolic fate, these separateisomers also have different binding affinities for the 5-HT4 receptor,thus suggesting different activities as well, and therefore differenttherapeutic uses. Thus, in a decreasing order of affinity for the 5-HT4receptor, the isomers can be ranked as follows (in parentheses are thebinding constant Ki values); compound IV (1.4 nM), compound VI (3.4 nM),compound III (28 nM), and compound V (72 nM). These binding experimentswere performed using the radiolabel displacement method described instandard textbooks and easily reproducible by persons skilled in the artof molecular biology.

As a conclusion to these considerations: when the 3 and 4 positions arecis relative to each other, compound I is a mixture of 4 isomers,consisting of 2 pairs of enantiomers. The first pair of enantiomers is(+)(R)-compound I and (−)(S)-compound I (compounds IV and V,respectively), the second pair of enantiomers is (−)(R)-compound I and(+)(S)-compound I (compounds III and VI, respectively). Within eachenantiomeric pair, each separate enantiomer has different propertiesregarding both their rate of hydrolysis by esterases and regarding theiraffinity at the 5-HT4 receptor. These different properties give themseparately advantageous therapeutic uses which are not interchangeable,i.e., which are specific to each isomer, and which are not applicable tothe racemic mixture. These differences of affinity at the receptor andthese differences in metabolic rates are not predictable and neither isit possible to dissect these properties when testing the racemicmixture.

Another aspect of the invention comprises a method for preparing acompound for formula II′

-   -   comprising converting a compound of formula (I′)    -   or its salt to a compound of formula (II′) or its salt,        respectively, wherein X₁ is a nitrogen protecting group and X₂        is selected from the group consisting of hydrogen and a nitrogen        protecting group (wherein commonly known and used N protecting        groups can be used, e.g., N-benzyl; N-nitrobenzyl; N-BoC;        N-oxide; N-paramethoxybenzyl; N-benzylsulfonyl) (preferably both        X₁ and X₂ are benzyl), and R is (C₁-C₈)alkyl, preferably        (C₁-C₄)alkyl, more preferably ethyl. In another embodiment, X₁        and X₂ are not both benzyl.

The invention also comprises compounds of formula II′

-   -   and salts thereof, wherein X₁ is a nitrogen protecting group and        X₂ is selected from the group consisting of hydrogen and a        nitrogen protecting group (wherein commonly known and used N        protecting groups can be used, e.g., N-benzyl; N-nitrobenzyl;        N-BoC; N-oxide; N-paramethoxybenzyl; N-benzylsulfonyl)        (preferably both X₁ and X₂ are benzyl), and R is (C₁-C₈)alkyl,        preferably (C₁-C₄)alkyl, more preferably ethyl. In another        embodiment, X₁ and X₂ are not both benzyl.

In another aspect, the invention comprises a method of making a compoundof formula III′

-   -   comprising treating the compound of formula (II′)    -   with an alkali metal hydroxide or hydride (e.g., NaOH, KOH,        sodium or potassium hydride, lithium aluminum hydride, etc.) to        yield the compound of formula (III′), wherein X₁ is a nitrogen        protecting group and X₂ is selected from the group consisting of        hydrogen and a nitrogen protecting group (wherein commonly known        and used N protecting groups can be used, e.g., N-benzyl;        N-nitrobenzyl; N-BoC; N-oxide; N-paramethoxybenzyl;        N-benzylsulfonyl) (preferably both X₁ and X₂ are benzyl), and R        is (C₁-C₈)alkyl, preferably (C₁-C₄)alkyl, more preferably ethyl.        In another embodiment, X₁ and X₂ are not both benzyl.

We have surprisingly found with experiments on

-   -   that using at least 12 equivalents of KOH and an 8-fold excess        of isopropyl alcohol under reflux in the hydrolysis reaction        results in virtually 100% conversion with virtually no        impurities. Using lesser amounts of KOH (e.g., 5 and 10 eq) gave        conversions only in the range of about 83-98% with impurities        ranging from 1.9% to 7.3%.

The invention also comprises a compounds of formula III′

and salts thereof, wherein X₁ is a nitrogen protecting group and X₂ isselected from the group consisting of hydrogen and a nitrogen protectinggroup (wherein commonly known and used N protecting groups can be used,e.g., N-benzyl; N-nitrobenzyl; N-BoC; N-oxide; N-paramethoxybenzyl;N-benzylsulfonyl) (preferably both X₁ and X₂ are benzyl). In anotherembodiment, X₁ and X₂ are not both benzyl.

In another aspect, the invention comprises a method of making a compoundof formula III″

-   -   comprising a) contacting a compound of formula III′    -   with a chiral resolving agent (e.g., tartaric acid, mandelic        acid, Mosher's acid, camphor sulphonic acid, etc.) to yield a        chiral salt of III″ and isolating the chiral salt of III″;    -   b) optionally recrystallizing the product of a); and    -   c) contacting the product of a) or b) with a base to yield the        compound for formula III″;    -   wherein X₁ is a nitrogen protecting group and X₂ is selected        from the group consisting of hydrogen and a nitrogen protecting        group (wherein commonly known and used N protecting groups can        be used, e.g., N-benzyl; N-nitrobenzyl; N-BoC; N-oxide;        N-paramethoxybenzyl; N-benzylsulfonyl) (preferably both X₁ and        X₂ are benzyl). In another embodiment, X₁ and X₂ are not both        benzyl.

Preferably in this aspect of the invention, the chiral resolving agentis (+)-2,3-dibenzoyl-D-tartaric acid and the chiral salt of III′ is a(3S,4R)-enantiomer (+)-2,3-dibenzoyl-D-tartrate salt.

It has been surprisingly found that the use of the chiral salt of(+)-2,3-dibenzoyl-D-tartaric acid, preferably in an amount of oneequivalent for two equivalents of compound of structure III′, leads toenhanced yield as compared to other chiral resolving agents. The use ofone equivalent of (+)-2,3-dibenzoyl-D-tartaric acid with two equivalentsof III′ results a yield of more than three times that obtained when oneequivalent of (+)-2,3-dibenzoyl-D-tartaric acid is contacted with twoequivalents of III′. Thus, in a preferred embodiment of making thecompound of formula III″, one equivalent of a chiral resolving agent(preferably (+)-2,3-dibenzoyl-D-tartaric acid) and two equivalents ofthe compound of formula III′ are used.

In another aspect, the invention comprises the compound of formula III″

and salts thereof, wherein X₁ is a nitrogen protecting group and X₂ isselected from the group consisting of hydrogen and a nitrogen protectinggroup (wherein commonly known and used N protecting groups can be used,e.g., N-benzyl; N-nitrobenzyl; N-BoC; N-oxide; N-paramethoxybenzyl;N-benzylsulfonyl) (preferably both X₁ and X₂ are benzyl). In anotherembodiment, X₁ and X₂ are not both benzyl.

In another aspect, the invention comprises a method of making a compoundof formula IV′

-   -   comprising contacting a compound for formula    -   with a (C₁-C₈)alkyl 6-halohexanoate (wherein the halo is        preferably bromo) to yield a compound of formula (IV′), wherein        R′ is (C₁-C₈)alkyl (preferably ethyl), and X₁ is a nitrogen        protecting group and X₂ is selected from the group consisting of        hydrogen and a nitrogen protecting group (wherein commonly known        and used N protecting groups can be used, e.g., N-benzyl;        N-nitrobenzyl; N-BoC; N-oxide; N-paramethoxybenzyl;        N-benzylsulfonyl) (preferably both X₁ and X₂ are benzyl). In        another embodiment, X₁ and X₂ are not both benzyl.

In another aspect, the invention a compound of formula IV′

and salts thereof, wherein R′ is (C₁-C₈)alkyl (preferably ethyl), and X₁is a nitrogen protecting group and X₂ is selected from the groupconsisting of hydrogen and a nitrogen protecting group (wherein commonlyknown and used N protecting groups can be used, e.g., N-benzyl;N-nitrobenzyl; N-BoC; N-oxide; N-paramethoxybenzyl; N-benzylsulfonyl)(preferably both X₁ and X₂ are benzyl). In another embodiment, X₁ and X₂are not both benzyl.

In another aspect, the invention comprises a method of making a compoundof formula V′

-   -   comprising contacting a compound of formula IV′    -   with (R)-quinuclidin-3-ol and a Lewis acid (e.g., a titanium        tetraalkoxide (e.g., Ti(OiPr)₄ (titanium tetraisopropoxide) and        Ti(OEt)₄ (titanium tetraethoxide)), TsOH (para toluenesulfonic        acid), K₂CO₃, and cat. DMAP (catalytic 4-dimethylaminopyridine))        in an organic solvent (e.g., toluene), wherein R′ is        (C₁-C₈)alkyl (preferably ethyl), and X₁ is a nitrogen protecting        group and X₂ is selected from the group consisting of hydrogen        and a nitrogen protecting group (wherein commonly known and used        N protecting groups can be used, e.g., N-benzyl; N-nitrobenzyl;        N-BoC; N-oxide; N-paramethoxybenzyl; N-benzylsulfonyl)        (preferably both X₁ and X₂ are benzyl). In another embodiment,        X₁ and X₂ are not both benzyl.

In another aspect, the invention comprises a compound of formula V′

and salts thereof, wherein X₁ is a nitrogen protecting group and X₂ isselected from the group consisting of hydrogen and a nitrogen protectinggroup (wherein commonly known and used N protecting groups can be used,e.g., N-benzyl; N-nitrobenzyl; N-BoC; N-oxide; N-paramethoxybenzyl;N-benzylsulfonyl) (preferably both X₁ and X₂ are benzyl). In anotherembodiment, X₁ and X₂ are not both benzyl.

In another aspect, the invention comprises a method of making a compoundof formula VI′

-   -   comprising removing groups X₁ and X₂ from a compound of formula        V′

wherein X₁ is a nitrogen protecting group and X₂ is selected from thegroup consisting of hydrogen and a nitrogen protecting group (whereincommonly known and used N protecting groups can be used, e.g., N-benzyl;N-nitrobenzyl; N-BoC; N-oxide; N-paramethoxybenzyl; N-benzylsulfonyl)(preferably both X₁ and X₂ are benzyl). In another embodiment, X₁ and X₂are not both benzyl.

In a preferred embodiment of this aspect of the invention, X₁ and X₂ arebenzyl and are removed by treatment of V′ with H₂/Pd/C or ammoniumformate/Pd are examples) to yield (R)-quinuclidin-3-yl6-[(3S,4R)-4-amino-3-methoxypiperidin-1-yl]hexanoate. We havesurprisingly found by hydrogenation with H₂/Pd/C this method providessignificant advantages over debenzylating methods using, for example,ammonium formate. Such methods are frequently messy and require silicagel column purification to remove reagents (e.g., ammonium formate),which is impractical for large scale production. Hydrogenation withH₂/Pd/C is extremely clean and does not require column purification.

In another aspect, the invention comprises a method of making(R)-quinuclidin-3-yl6-((3S,4R)-4-(4-amino-5-chloro-2-methoxybenzamido)-3-methoxypiperidin-1-yl)hexanoate(VII′) comprising contacting a compound of formula VI′

with 4-amino-5-chloro-2-methoxybenzoic acid. Preferably said contactingis with EDCI (1-ethyl-3-(3′-dimethylaminopropyl)carbodiimide), HOBt(1-hydroxbenzotriazole), HOSU (N-hydroxysuccinimide), HONB(N-hydroxy-5-norbene-endo-2,3-dicarboxamide), isobutyl chloroformate,pivaloyl chloride, or DCC (dicyclohexylcarbodiimide). Most preferablysaid contacting is with a pivaloyl halide (preferably pivaloylchloride). It was unexpectedly found that the use of pivaloyl chloridegave a significantly cleaner reaction profile and the product was mucheasier to purify compared to other acylating agents. The result is ahigher yield and significantly greater purity of the compound comparedto methods employing other acylating agents.

Compounds of formulae I′, II′, III′, III″, IV′, V′, and VI′ are alluseful intermediates in the production of (R)-quinuclidin-3-yl6-((3S,4R)-4-(4-amino-5-chloro-2-methoxybenzamido)-3-methoxypiperidin-1-yl)hexanoate.

In another aspect, the invention comprises combinations of the foregoingmethods. As used herein, a method described as X—Y is a methodcomprising the combination of making a compound of formula “X” followedby a method of making a compound of formula “Y” and, similarly X—Y-Z isthe method X—Y followed by the method of making the compound of formula“Z,” etc. Accordingly, this aspect of the invention includes, withoutlimitation, methods I′-II′, II′-III′, III′-III″, III″-IV′, IV′-V′,V′-VI′, VI′-VII′, I′-II′-III′, II′-III′-III″, III′-III″-IV′,III″-IV′-V′, IV′-V′-VI′, V′-VI′-VII′, I′-II′-III′-III″,II′-III′-III″-IV′, III′-III″-IV′-V′, III″-IV′-V′-VI′, andIV′-V′-VI′-VII′.

Another aspect of the invention comprises a method for preparing(R)-quinuclidin-3-yl6-((3S,4R)-4-(4-amino-5-chloro-2-methoxybenzamido)-3-methoxypiperidin-1-yl)hexanoateor a salt thereof, comprising:

-   -   1) optionally converting a compound of formula (I′)    -   to a salt, wherein R is (C₁-C₈)alkyl (preferably (C₁-C₆)alkyl,        (C₁-C₄)alkyl, or ethyl);    -   2) converting the compound of formula (I′) or its salt to a        compound of formula (II′)    -   or its salt, respectively, wherein X₁ is a nitrogen protecting        group and X₂ is selected from the group consisting of hydrogen        and a nitrogen protecting group (wherein commonly known and used        N protecting groups can be used, e.g., N-benzyl; N-nitrobenzyl;        N-BoC; N-oxide; N-paramethoxybenzyl; N-benzylsulfonyl)        (preferably both X₁ and X₂ are benzyl);    -   3) treating the compound of formula (II′) with an alkali metal        hydroxide or hydride (e.g., NaOH, KOH, sodium or potassium        hydride, lithium aluminum hydride, etc.) to yield a compound of        formula (III′)    -   4) producing a chiral salt of III′ by treating the compound of        formula (III″) with a chiral resolving agent (e.g., tartaric        acid, mandelic acid, Mosher's acid, camphor sulphonic acid,        etc., or, preferably, (+)-2,3-dibenzoyl-D-tartaric acid to yield        a chiral salt (e.g., (3S,4R)-enantiomer        (+)-2,3-dibenzoyl-D-tartrate salt when        (+)-2,3-dibenzoyl-D-tartaric acid is the chiral resolving        agent)) and isolating the cis isomer of III′ thereby produced;    -   5) optionally recrystallizing the product of 4;    -   6) basifying the product of 4 or 5 to yield the free base form        of the product of 4 or 5;    -   7) contacting the product of 6 with a (C₁-C₈)alkyl        6-halohexanoate (wherein the halo is preferably bromo) to yield        a compound of formula (IV′)    -   wherein R′ is (C₁-C₈)alkyl (preferably ethyl);    -   8) treating the product of 7 with (R)-quinuclidin-3-ol and a        Lewis acid (e.g., a titanium tetraalkoxide (e.g., Ti(OiPr)₄        (titanium tetraisopropoxide) and Ti(OEt)₄ (titanium        tetraethoxide)), TsOH (para toluenesulfonic acid), K₂CO₃, and        cat. DMAP (catalytic 4-dimethylaminopyridine)) in an organic        solvent (e.g., toluene) to yield a compound of formula (V′)    -   9) deprotecting the 4-amino group of the product of 8 (e.g.,        with H₂/Pd/C or ammonium formate/Pd are examples) to yield        (R)-quinuclidin-3-yl        6-[(3S,4R)-4-amino-3-methoxypiperidin-1-yl]hexanoate;    -   10) acylating the product of 9 with        4-amino-5-chloro-2-methoxybenzoic acid (e.g., with EDCI        (1-ethyl-3-(3′-dimethylaminopropyl)carbodiimide); HOBt        (1-hydroxbenzotriazole); HOSU (N-hydroxysuccinimide); HONB        (N-hydroxy-5-norbene-endo-2,3-dicarboxamide); isobutyl        chloroformate; pivaloyl chloride; DCC        (dicyclohexylcarbodiimide)) to yield (R)-quinuclidin-3-yl        6-((3S,4R)-4-(4-amino-5-chloro-2-methoxybenzamido)-3-methoxypiperidin-1-yl)hexanoate;    -   11) optionally converting the product of 10 to a salt.

A further aspect of the invention is process for preparing(R)-quinuclidin-3-yl6-((3S,4R)-4-(4-amino-5-chloro-2-methoxybenzamido)-3-methoxypiperidin-1-yl)hexanoateor a salt thereof, comprising:

-   -   1) converting a compound that is ethyl        4-amino-3-methoxypiperidine-1-carboxylate    -   to a salt;    -   2) converting the ethyl        4-amino-3-methoxypiperidine-1-carboxylate salt to ethyl        4-(diphenylamino)-3-methoxypiperidine-1-carboxylate    -   3) treating the ethyl        4-(diphenylamino)-3-methoxypiperidine-1-carboxylate with an        alkali metal hydroxide or hydride to yield        3-methoxy-N,N-diphenylpiperidin-4-amine    -   4) producing a chiral salt of        3-methoxy-N,N-diphenylpiperidin-4-amine by treating the        3-methoxy-N,N-diphenylpiperidin-4-amine with a chiral resolving        agent and isolating the cis isomer of the salt of        3-methoxy-N,N-diphenylpiperidin-4-amine thereby produced;    -   5) optionally recrystallizing the product of 4;    -   6) basifying the product of 4 or 5 to yield the free base form        of the product of 4 or 5    -   7) alkylating the product of 6 with ethyl 6-bromohexanoate to        yield ethyl        6-((3S,4R)-4-(diphenylamino)-3-methoxypiperidin-1-yl)hexanoate    -   8) esterifying the ethyl        6-((3S,4R)-4-(diphenylamino)-3-methoxypiperidin-1-yl)hexanoate        with (R)-quinuclidin-3-ol to yield (R)-quinuclidin-3-yl        6-((3S,4R)-4-(diphenylamino)-3-methoxypiperidin-1-yl)hexanoate    -   9) deprotecting the 4-amino group of the product of 8 to yield        (R)-quinuclidin-3-yl        6-[(3S,4R)-4-amino-3-methoxypiperidin-1-yl]hexanoate;    -   10) acylating the product of 9 with        4-amino-5-chloro-2-methoxybenzoic acid to yield        (R)-quinuclidin-3-yl        6-((3S,4R)-4-(4-amino-5-chloro-2-methoxybenzamido)-3-methoxypiperidin-1-yl)hexanoate;    -   11) optionally converting the product of 10 to a salt.

Preferably in this aspect of the invention the salt of step 1 is HCl.

Preferably in this aspect of the invention the alkali metal hydroxide ofstep 3 is KOH.

Preferably in this aspect of the invention the chiral salt of step 4 is(+)-2,3-dibenzoyl-D-tartaric acid, which, upon reaction with III′,yields a (3S,4R)-enantiomer (+)-2,3-dibenzoyl-D-tartrate salt.

Preferably in this aspect of the invention the reaction conditions ofstep 8 comprise Ti(OiP)₄ (titanium (IV) isopropoxide).

Preferably in this aspect of the invention the reaction conditions ofstep 9 comprise H₂/Pd/C.

Preferably in this aspect of the invention the reaction conditions ofstep 10 comprise pivaloyl chloride.

Definitions

As used herein, the term “alkyl” includes those alkyl groups of adesigned number of carbon atoms. Alkyl groups may be straight, orbranched. Examples of “alkyl” include methyl, ethyl, propyl, isopropyl,butyl, iso-, sec- and tert-butyl, pentyl, hexyl, heptyl, 3-ethylbutyl,and the like. If the number of carbon atoms is not specified, thesubject “alkyl” moiety has from 1 to 6 carbons.

The term “alkoxy” represents an alkyl group of indicated number ofcarbon atoms attached to the parent molecular moiety through an oxygenbridge. Examples of alkoxy groups include, for example, methoxy, ethoxy,propoxy and isopropoxy.

By “aryl” is meant an aromatic carbocyclic group having a single ring(e.g., phenyl) that is optionally fused or otherwise attached to otheraromatic hydrocarbon rings or non-aromatic hydrocarbon rings. “Aryl”includes multiple condensed rings in which at least one is aromatic,(e.g., 1,2,3,4-tetrahydronaphthyl, naphthyl), wherein each ring isoptionally mono-, di-, or trisubstituted with the groups identifiedbelow, as well as multiple rings that are not fused, such as, forexample, biphenyl or binaphthyl. Preferred aryl groups of the presentinvention are phenyl, 1-naphthyl, 2-naphthyl, indanyl, indenyl,dihydronaphthyl, fluorenyl, tetralinyl or6,7,8,9-tetrahydro-5H-benzo[a]cycloheptenyl. More preferred are phenyl,biphenyl, and naphthyl. Most preferred is phenyl. The aryl groups hereinare unsubstituted or, as specified, substituted in one or moresubstitutable positions with various groups. For example, such arylgroups may be optionally substituted with, for example, C₁-C₆ alkyl,C₁-C₆ alkoxy, halogen, hydroxy, cyano, nitro, amino,mono(C₁-C₆)alkylamino, di(C₁-C₆)alkylamino, C₂-C₆alkenyl, C₂-C₆alkynyl,C₁-C₆ haloalkyl, C₁-C₆ haloalkoxy, amino(C₁-C₆)alkyl,mono(C₁-C₆)alkylamino(C₁-C₆)alkyl or di(C₁-C₆)alkylamino(C₁-C₆)alkyl.

The term “haloalkoxy” refers to an alkoxy group substituted with atleast one halogen atom and optionally further substituted with at leastone additional halogen atom, where each halogen is independently F, Cl,Br or I. Preferred halogens are F or Cl. Preferred haloalkoxy groupscontain 1-6 carbons, more preferably 1-4 carbons, and still morepreferably 1-2 carbons. “Haloalkoxy” includes perhaloalkoxy groups, suchas OCF₃ or OCF₂CF₃.

The term “heteroaryl” refers to an aromatic ring system containing atleast one heteroatom selected from nitrogen, oxygen, and sulfur. Theheteroaryl ring may be fused or otherwise attached to one or moreheteroaryl rings, aromatic or non-aromatic hydrocarbon rings orheterocycloalkyl rings. Examples of heteroaryl groups include, forexample, pyridyl, pyrimidinyl, quinolinyl, benzothienyl, indolyl,indolinyl, pyridazinyl, pyrazinyl, isoindolyl, isoquinolyl,quinazolinyl, quinoxalinyl, phthalazinyl, imidazolyl, isoxazolyl,pyrazolyl, oxazolyl, thiazolyl, indolizinyl, indazolyl, benzothiazolyl,benzimidazolyl, benzofuranyl, furanyl, thienyl, pyrrolyl, oxadiazolyl,thiadiazolyl, benzo[1,4]oxazinyl, triazolyl, tetrazolyl, isothiazolyl,naphthyridinyl, isochromanyl, chromanyl, tetrahydroisoquinolinyl,isoindolinyl, isobenzotetrahydrofuranyl, isobenzotetrahydrothienyl,isobenzothienyl, benzoxazolyl, pyridopyridinyl, benzotetrahydrofuranyl,benzotetrahydrothienyl, purinyl, benzodioxolyl, triazinyl, pteridinyl,benzothiazolyl, imidazopyridinyl, imidazothiazolyl,dihydrobenzisoxazinyl, benzisoxazinyl, benzoxazinyl,dihydrobenzisothiazinyl, benzopyranyl, benzothiopyranyl, chromonyl,chromanonyl, pyridinyl-N-oxide, tetrahydroquinolinyl, dihydroquinolinyl,dihydroquinolinonyl, dihydroisoquinolinonyl, dihydrocoumarinyl,dihydroisocoumarinyl, isoindolinonyl, benzodioxanyl, benzoxazolinonyl,pyrrolyl N-oxide,, pyrimidinyl N-oxide, pyridazinyl N-oxide, pyrazinylN-oxide, quinolinyl N-oxide, indolyl N-oxide, indolinyl N-oxide,isoquinolyl N-oxide, quinazolinyl N-oxide, quinoxalinyl N-oxide,phthalazinyl N-oxide, imidazolyl N-oxide, isoxazolyl N-oxide, oxazolylN-oxide, thiazolyl N-oxide, indolizinyl N-oxide, indazolyl N-oxide,benzothiazolyl N-oxide, benzimidazolyl N-oxide, pyrrolyl N-oxide,oxadiazolyl N-oxide, thiadiazolyl N-oxide, triazolyl N-oxide, tetrazolylN-oxide, benzothiopyranyl S-oxide, benzothiopyranyl S,S-dioxide.Preferred heteroaryl groups include pyridyl, pyrimidyl, quinolinyl,indolyl, pyrrolyl, furanyl, thienyl, and imidazolyl. More preferredheteroaryl groups include pyridyl, pyrrolyl, and indolyl. The heteroarylgroups herein are unsubstituted or, as specified, substituted in one ormore substitutable positions with various groups. For example, suchheteroaryl groups may be optionally substituted with, for example, C₁-C₆alkyl, C₁-C₆ alkoxy, halogen, hydroxy, cyano, nitro, amino,mono(C₁-C₆)alkylamino, di(C₁-C₆)alkylamino, C₂-C₆alkenyl, C₂-C₆alkynyl,C₁-C₆ haloalkyl, C₁-C₆ haloalkoxy, amino(C₁-C₆)alkyl,mono(C₁-C₆)alkylamino(C₁-C₆)alkyl or di(C₁-C₆)alkylamino(C₁-C₆)alkyl.

The term “heterocycloalkyl” refers to a ring or ring system containingat least one heteroatom that is preferably selected from nitrogen,oxygen, and sulfur, wherein said heteroatom is in a non-aromatic ring.The heterocycloalkyl ring is optionally fused to or otherwise attachedto other heterocycloalkyl rings and/or non-aromatic hydrocarbon ringsand/or phenyl rings. Preferred heterocycloalkyl groups have from 3 to 7members. More preferred heterocycloalkyl groups have 5 or 6 members.Examples of heterocycloalkyl groups include, for example,aza-bicyclo[2.2.2]octyl, aza-bicyclo[3.2.1]octyl, morpholinyl,thiomorpholinyl, thiomorpholinyl S-oxide, thiomorpholinyl S,S-dioxide,piperazinyl, homopiperazinyl, pyrrolidinyl, pyrrolinyl,tetrahydropyranyl, piperidinyl, tetrahydrofuranyl, tetrahydrothienyl,homopiperidinyl, homomorpholinyl, homothiomorpholinyl,homothiomorpholinyl S,S-dioxide, oxazolidinonyl, dihydropyrazolyl,dihydropyrrolyl, dihydropyrazinyl, dihydropyridinyl, dihydropyrimidinyl,dihydrofuryl, dihydropyranyl, tetrahydrothienyl S-oxide,tetrahydrothienyl S,S-dioxide and homothiomorpholinyl S-oxide. Preferredheterocycloalkyl groups include aza-bicyclo[2.2.2]octyl,aza-bicyclo[3.2.1]octyl, piperidinyl, piperazinyl, pyrrolidinyl,thiomorpholinyl, S,S-dioxothiomorpholinyl, morpholinyl, andimidazolidinyl. More preferred are aza-bicyclo[2.2.2]octyl,aza-bicyclo[3.2.1]octyl, piperidinyl, piperazinyl, pyrrolidinyl,imidazolidinyl, and morpholinyl. The heterocycle groups herein areunsubstituted or, as specified, substituted in one or more substitutablepositions with various groups. For example, such heterocycle groups maybe optionally substituted with, for example, C₁-C₆ alkyl, C₁-C₆ alkoxy,halogen, hydroxy, cyano, nitro, amino, mono(C₁-C₆)alkylamino,di(C₁-C₆)alkylamino, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆ haloalkyl,C₁-C₆ haloalkoxy, amino(C₁-C₆)alkyl, mono(C₁-C₆)alkylamino(C₁-C₆)alkyl,di(C₁-C₆)alkylamino(C₁-C₆)alkyl or ═O.

The term “pharmaceutically acceptable salts” or “a pharmaceuticallyacceptable salt thereof” refer to salts prepared from pharmaceuticallyacceptable non-toxic acids or bases including inorganic acids and basesand organic acids and bases. Since the compound of the present inventionis basic, salts may be prepared from pharmaceutically acceptablenon-toxic acids. Suitable pharmaceutically acceptable acid additionsalts for the compound of the present invention include acetic,benzenesulfonic (besylate), benzoic, camphorsulfonic, citric,ethenesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric,isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic,nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric,p-toluenesulfonic, and the like. Preferred acid addition salts are thechloride and sulfate salts. In the most preferred aspect, structuraland/or functional analogs of cisapride are administered as the free baseor as the mono or dihydrochloride salt.

As used herein, the terms “treatment” and “treating” encompassprophylactic administration of the compound or a pharmaceuticalcomposition comprising the compound (“prophylaxis”) as well as remedialtherapy to reduce or eliminate a disease or disorder mentioned herein.Prophylactic administration is intended for prevention of disorders andmay be used to treat a subject that is at risk of having or sufferingfrom one or more disorders mentioned herein. Thus, as used herein, theterm “treatment”, or a derivative thereof, contemplates partial orcomplete inhibition of the stated disease state, when an activeingredient of the invention is administered prophylactically orfollowing the onset of the disease state for which such activeingredient of the is administered. “Prophylaxis” refers toadministration of the active ingredient(s) to a mammal to protect themammal from any of the disorders set forth herein, as well as others.

The term “therapeutically effective amount” refers to an amountnecessary to achieve a derived therapeutic effect such as: 1) an amountsufficient to alleviate reflux disease, 2) an amount sufficient toalleviate nausea and vomiting, or 3) an amount sufficient to alleviate acondition caused by gastrointestinal motility dysfunction.Therapeutically effective amounts of structural and/or functionalanalogs of cisapride are encompassed by the above-described dosageamounts and dose frequency schedule.

A “mammal” may be, for example, a mouse, rat, pig, horse, rabbit, goat,cow, cat, dog, or human. In a preferred aspect, the mammal is a human.

The term “individual(s)” is defined as a single mammal to which isadministered a compound of the present invention. The mammal may be, forexample, a mouse, rat, pig, horse, rabbit, goat, cow, cat, dog, orhuman. In a preferred aspect, the individual is a human.

The term “esterified cisapride” means therapeutic compounds of thesubject invention that are structural and/or functional analogs ofcisapride, which contain a hydrolysable group, generally an ester, thatdoes not detract from the ability of these compounds to provide atherapeutic benefit, but which makes these compounds more susceptible todegradation by hydrolases, particularly serum and/or cytosolicesterases, and which reduces the interaction of the cytochrome P-450drug detoxification system with the cisapride compounds.Esterase-mediated metabolism of esterified cisapride compounds reducesthe role of the cytochrome P-450 drug detoxification system in cisapridemetabolism and reduces or eliminates adverse effects caused bycisapride.

The term “structural analog” as used herein means that a describedcompound shares structural characteristics with a parent compound. Forexample, a structural analog of cisapride may share one or morestructural characteristics with the parent cisapride compound, such as asubstituted aryl ring connected to a piperdine ring through an amidelinker, but differ structurally in other ways, such as the inclusion ordeletion of one or more other chemical moieties.

The term “functional analog” as used herein means that a describedcompound shares a functional characteristic with a parent compound. Forexample, a functional analog of cisapride may share few, if any,structural characteristics with cisapride, but affect a similarfunction, for example, 5-HT₄ agonism.

The term “adverse effects” includes, but is not limited to,gastrointestinal disorders such as diarrhea, abdominal cramping, andabdominal grumbling; tiredness; headache; increased systolic pressure;death; ventricular tachycardia; ventricular fibrillation; torsades depointes; QT prolongation; increased heart rate; neurological and CNSdisorders; and interaction of cisapride with other drugs givenconcurrently such as but not limited to digoxin, diazepam, ethanol,acenocoumarol, cimetidine, ranitidine, paracetamol, and propranolol.

The term “gastroesophageal reflux disease” as used herein means theincidence of, and the symptoms of, those conditions causing the backwardflow of the stomach contents into the esophagus.

The terms “eliciting an anti-emetic effect” and “anti-emetic therapy” asused herein mean providing relief from or preventing the symptoms ofnausea and vomiting induced spontaneously or associated with emetogeniccancer chemotherapy or irradiation therapy.

The term “treating a condition caused by gastrointestinal motilitydysfunction” as used herein means treating the symptoms and conditionsassociated with this disorder which include, but are not limited to,gastroesophageal reflux disease, dyspepsia, gastroparesis, constipation,post-operative ileus, and intestinal pseudo-obstruction.

The term “prokinetic” as used herein means the enhancement ofperistalsis in, and thus the movement through the gastrointestinaltract.

The term “dyspepsia” as used herein means a condition characterized byan impairment of the power or function of digestion that can arise as asymptom of a primary gastrointestinal dysfunction or as a complicationdue to other disorders such as appendicitis, gallbladder disturbances,or malnutrition.

The term “gastroparesis” as used herein means a paralysis of the stomachbrought about by a motor abnormality in the stomach or as a complicationof diseases such as diabetes, progressive systemic sclerosis, anorexianervosa, or myotonic dystrophy.

The term “constipation” as used herein means a condition characterizedby infrequent or difficult evacuation of feces resulting from conditionssuch as lack of intestinal muscle tone or intestinal spasticity.

The term “post-operative ileus” as used herein means an obstruction inthe intestine due to a disruption in muscle tone following surgery.

The term “intestinal pseudo-obstruction” as used herein means acondition characterized by constipation, colicky pain, and vomiting, butwithout evidence of physical obstruction.

Preparation of Compounds

While the chemical synthesis of various analogs of cisapride can beperformed by the methods described in European Patent Application No.0,076,530 A2 published Apr. 13, 1983, WO 01/093849, U.S. Pat. Nos.4,962,115 and 5,057,525 and in Van Daele et al., Drug Development Res.8: 225-232 (1986), the disclosures of which are incorporated herein byreference in their entireties, compounds of the invention are preferablymade according to the disclosure of Methods 3-6.

The invention is illustrated further by some examples, which follow themethods. The methods and examples are not to be construed as limitingthe invention in scope or spirit to the specific procedures described inthem. Those having skill in the art will recognize that the startingmaterials may be varied and additional steps employed to producecompounds encompassed by the invention, as demonstrated by the followingexamples. Those skilled in the art will also recognize that it may benecessary to utilize different solvents or reagents to achieve some ofthe above transformations. In some cases, protection of reactivefunctionalities may be necessary to achieve the above transformations.In general, such need for protecting groups, as well as the conditionsnecessary to attach and remove such groups, will be apparent to thoseskilled in the art of organic synthesis. When a protecting group isemployed, deprotection step may be required. Suitable protecting groupsand methodology for protection and deprotection such as those describedin Protecting Groups in Organic Synthesis by T. Greene are well knownand appreciated in the art.

Unless otherwise specified, all reagents and solvents are of standardcommercial grade and are used without further purification. Theappropriate atmosphere to run the reaction under, for example, air,nitrogen, hydrogen, argon and the like, will be apparent to thoseskilled in the art.

Method 1 Preparation of (R)-quinuclidin-3-yl6-((3S,4R)-4-(4-amino-2-chloro-6-methoxybenzamido)-3-methoxypiperidin-1-yl)hexanoate

Step 1: Synthesis of ethyl4-(dibenzylamino)-3-methoxypiperidine-1-carboxylate (1):

To a solution of racemic ethyl 4-amino-3-methoxypiperidine-1-carboxylate(1 part by mole) in DMF were added benzyl bromide (about 2.2 part bymole), potassium carbonate (about 2.4 part by mole) and potassium iodide(about 0.2 part by mole) respectively. The reaction was heated to about80° C. (in the specification, delta, or “A,” refers to heat). Afterabout 6 hours, the reaction was slowly diluted with water (about 12parts by volume) and extracted with, for example, ethyl acetate. Theorganic layer was washed with brine and then dried over anhyh. Na₂SO₄.Subsequent filtration and concentration of the solvent provided the 1 asthe yellow-orange oil (1 part by mole).

Step 2. Synthesis of N,N-dibenzyl-3-methoxypiperidin-4-amine (2):

To a solution of 1 was added NaOH (about 10 part by mole) in isopropanoland the mixture was stirred and heated to reflux. After about 3 to about5 hours, the reaction was cooled to room temperature and the alcoholicsolvent was removed via rotary evaporation. The mixture was diluted withwater and extracted with ethyl acetate. The organic layer was brinedwashed before drying over anhyh. Na₂SO₄. Subsequent filtration andconcentration of the solvent provided a crude oil which was purifiedover SiO₂ (CH₂Cl₂:MeOH:NH₄OH; (about) 15:1:0.01) to furnish 2.

Step 3. Synthesis of (3S,4R)-N,N-dibenzyl-3-methoxypiperidin-4-amine(3):

(+)-2,3-Dibenzoyl-D-tartaric acid (about 1.2 part by weight) isdissolved in ethanol before slowly adding to a solution of 2 (about 1part by weight). The solution is gently warmed and then allowed to coolto room temperature to crystallize the salt product. The salt isfiltered and washed with EtOH/H₂O before suspending in water andbasifying by adding aq. NaOH (7%, wt/wt) until the pH reaches about 12.The suspension is stirred vigorously at rt and the solid is filteredaway, washed with water and vacuum dried to furnish the cis-isomer 3.

Step 4. Synthesis of ethyl6-((3S,4R)-4-(dibenzylamino)-3-methoxypiperidin-1-yl)hexanoate (4):

To a solution of 3 (1 part by mole) in DMF are added ethylbromohexanoate (about 1.2 part by mole), potassium carbonate (about 1.4part by mole) and potassium iodide (about 0.2 part by mole)respectively. The reaction is then heated to 80° C. After about 8 h, thereaction is slowly diluted with water (about 12 part by volume) andextracted with ethyl acetate. The organic layer is washed with brine andthen dried over anhyd. Na₂SO₄. Subsequent filtration and concentrationof the solvent furnishes the crude material. Purification over SiO₂ andgives the alkylated material 4.

Step 5. Synthesis of (R)-quinuclidin-3-yl6-((3S,4R)-4-(dibenzylamino)-3-methoxypiperidin-1-yl)hexanoate (5):

Titanium tetraethoxide is added to a mixture of 4 (1 part by mole) and(R)-(−)-3-quinuclidinol (1 part by mole) in toluene. The reactionmixture is equipped with a dean-stark apparatus before heating to about90° C. and partial vacuum is then applied (additional toluene is addedas needed to main the requisite solvent level). The mixture is thencooled to rt and the reaction is diluted with ethyl acetate and thenwater is added to the resulting mixture. The organic layer is separated,brine washed, dried over anhyd. Na₂SO₄, filtered and concentrated.Purification over Sio₂ gives the enantiomerically enriched 5.

Step 6. Synthesis of (R)-quinuclidin-3-yl6-((3S,4R)-4-amino-3-methoxypiperidin-1-yl)hexanoate (6):

A solution of 5 (1 part by mole) in EtOH is added to a reaction flaskcontaining palladium on carbon (about 0.2 part by mole). The mixture isthen evacuated of air before subjecting to hydrogenolysis condition byusing atmospheric H₂. Upon completion of the reaction, the palladium isfiltered off under a pad of celite followed by EtOH washes. Thefiltrated is concentrated via rotary evaporation to furnish 6.

Step 7. Synthesis of (R)-quinuclidin-3-yl6-((3S,4R)-4-(4-amino-2-chloro-6-methoxybenzamido)-3-methoxypiperidin-1-yl)hexanoate(7):

To a solution of, for example, ethyl chloroformate (1 part by mole) inTHF at about 0° C. is added the benzoic acid (1 part by mole) inportions. The mixture is warmed to rt for about 1 h before cooling toabout 0° C. and adding dropwise a solution of 6 (1 part by mole). Thereaction is then warmed to rt. Upon completion of the reaction, reactionis quenched by addition of a sat'd solution of NaHCO₃ and extractingover EA. The organic layer is brine washed, dried over anhyd. Na₂SO₄,filtered and concentrated to furnish the desired product 7.

Method 2

Synthesis of (R)-quinuclidin-3-yl6-((3S,4R)-4-(4-amino-5-chloro-2-methoxybenzamido)-3-methoxypiperidin-1-yl)hexanoate(or6-[4R-(4-amino-5-chloro-2-methoxy-benzoylamino)-3S-methoxy-piperidin-1-yl]-hexanoicacid 1-aza-bicyclo[2.2.2]oct-3′R-yl ester; ATI-7505):

Under acidic conditions, 1-benzylpiperidin-4-one (1) and hydrobromicacid are reacted in the presence of acetic acid to generateN-benzyl-3-bromopiperidin-4-one (2). Treatment of 2 with a sodiummethoxide and methanol solution provides1-benzyl-4,4-dimethoxypiperidin-3-ol (3). [The presence of thebeta-amino group negates the possibility of a Favorskii-type reaction.]Methylation of the hydroxyl group is done using a hydride base followedby treatment with iodomethane in the presence of DMF as the solvent tofurnish compound 4.

Subsequent acetal hydrolysis using 1% sulfuric acid in the presence ofheat yields a piperidine 5, which can then undergo a reductive aminationusing, for example, sodium cyanoborohydride and ammonium acetate inmethanol to yield 1-benzyl-3-methoxypiperidin-4-amine (6). At thisstage, 6 can undergo a chiral resolution technique. This can beaccomplished, for example, using (+)-DBT or other variant of tartaricacid in the presence of the suitable solvent to afford exclusivelyasymmetrically pure compound 7. Boc group protection of the primaryamine in 7 can be accomplished using Boc anhydride in the presence ofTHF solvent to obtain 8. A debenzylation reaction by hydrogenolysisusing Pd/C in methanol in the presence of atmospheric hydrogen gas setthe stage for the alkylation step. Treatment of 6-bromohexanenitrile inthe presence of mild base and DMF generates compound 10. A nitrile toester conversion using (R)-quinuclidinol in the presence of dilute acidgenerates 11. Subsequent removal of the Boc group using TFA furnishesthe free amine, which can undergo a coupling reaction with requisitebenzoic acid in the presence of a coupling reagent such as ethylchloroformate (and more preferably isobutyl chloroformate) to affordATI-7505 as an enantiomerically pure material.

Alternatively, compound 9 can be alkylated using ethyl 6-bromohexanoatein the presence of mild base. Subsequent removal of the Boc group yieldscompound 14. Titanium mediated transesterification of 14 using(R)-quinuclidinol and titanium tetraethoxide in toluene solventgenerates ATI-7505. Carlsburg esterase hydrolyzes esters that are of theS-configuration, therefore leaving intact esters that are of the Rconfiguration. Therefore treatment of diasteriomeric mixtures of 15 withthe Carlsburg esterase may also yield ATI-7505.

Method 3

Alternate synthesis of (R)-quinuclidin-3-yl6-((3S,4R)-4-(4-amino-5-chloro-2-methoxybenzamido)-3-methoxypiperidin-1-yl)hexanoatedihydrochloride salt—ATI-7505 dihydrochloride salt:

With vigorous stirring hydrogen chloride in diethyl ether (about 1.4parts by mole) was slowly added to a solution of piperidine carbamate(about 1.0 part by mole). The mixture was allowed to stir for about 8hours before filtering and washing with diethyl ether. The white solidwas further washed with dichloromethane and diethyl ether (about 1.1ratio by volume) to remove impurities and was subsequently dried undervacuum to obtain the racemic piperidine carbamate hydrochloride salt asa white solid.

Benzyl bromide (about 2.2 parts by mole) was added to a mixture of thepiperidine hydrochloride (about 1.0 parts by mole), potassium carbonate(K₂CO₃, about 2.4 parts by mole), and potassium iodide (KI, about 0.1parts by mole) in dimethylformamide at room temperature (rt). Thereaction mixture was heated to about 75° C. After about 18 hours, thereaction was cooled to rt, diluted with water and extracted with ethylacetate (EA). The organic layer was washed with brine and then driedover anhyd. (anhydrous) sodium sulfate (Na₂SO₄). Subsequent filtrationunder vacuum and concentration provided the crude oil product. Theproduct was precipitated out by adding a mixture of isopropanol andwater (about 1:1 volume ratio) and with stirring. Following vacuumfiltration provided a dibenzylamino piperidine as a white solid.

Potassium hydroxide (about 10 parts by mole) was added in portions to astirred solution of dibenzylamino carbamate (about 1.0 parts by mole) inisopropanol at room temperature and the mixture was stirred and heatedto reflux. After about 5 hours the reaction was cooled to roomtemperature and the solvent was removed under vacuum to approximatelyhalf volume. The reaction mixture was diluted with water and extractedwith ethyl acetate. Following brine wash, the product was dried overanhydrous Na₂SO₄. Subsequent vacuum filtration provided a piperidine asa semisolid.

Chiral resolution of 3,4-disubstituted piperidine:

(+)-2,3-Dibenzoyl-D-tartaric acid [(+)-DBT; about 1.0 parts by mole] wasdissolved in methanol and was added slowly to a heated solution (about70° C.) of disubstituted piperidine (about 1.0 parts by mole) inmethanol and water (about 1:1 ratio by volume). The mixture was stirredat this temperature for about 1 hour before removing the heat andallowing it to stir at room temperature for several hours, e.g., about16 hours in one case. The product salt was collected by vacuumfiltration and rinsed with methanol and water (about 1:1 ratio byvolume). The wet-cake was collected and recrystallized two more timesusing the same procedure as above.

The wetcake was suspended in water and 1N sodium hydroxide was added toit (to a pH of about 12). The resulting suspension was stirred for about3 hours at room temperature before extracting with ethyl acetate. Theorganic layer was washed with brine, filtered and concentrated toprovide the enantiomerically enriched 3,4-disubstituted piperidineproduct as a white solid.

To a mixture of the piperidine (about 1.0 parts by mole), K₂CO₃ (about1.2 parts by mole) and KI (about 0.1 parts by mole) in DMF solvent wasslowly added ethyl 6-bromohexanoate (about 1.1 parts by mole). Thereaction was stirred-heat at about 70° C. for about 10 hours beforecooling to room temperature and diluting with water and extracting withethyl acetate. The organic layer was separated and then washed withbrined and finally dried over anhydrous Na₂SO₄. Subsequent filtrationand concentration provided the crude oil. The crude product was purifiedvia flash column chromatography (e.g., 1:1 ratio of hexanes:ethylacetate by volume) to provide the product as a light brownish oil.

To a mixture of the above piperidine ester (about 1.0 parts by mole) and(3R)-quinuclidinol (about 4.0 parts by mole) was added titanium(IV)tetraethoxide (about 1.0 parts by mole) at room temperature. Thereaction was heated to about 85° C. and was run under partial pressureto remove any evolving ethanol. After about 18 hours, the reaction wascooled to rt before diluting with ethyl acetate and quenching withwater. The organic layers were then washed with brine and dried overanhydrous Na₂SO₄. Following concentration, the crude oil was purifiedvia flash column chromatography (e.g., about 100:10:1;CH₂Cl₂:MeOH:NH₄OH) to provide the product as a clear oil.

To a reaction flask containing palladium on carbon was added a solutionof the above dibenzyl piperidine ester (about 1.0 parts by mole) inmethanol, and to this mixture was added ammonium formate (about 4 partsby mole). The reaction was heated to reflux and after about 10 hours,the reaction flask was cooled to room temperature and the palladium oncarbon was filtered away, e.g., through a pad of celite. The filtratewas concentrated to an oil, which was purified via flash columnchromatography (e.g., SiO₂: about 150:10:1; CH₂Cl₂:CH₃OH:NH₄)H) toprovide the amino piperidine ester product as a yellow oil.

To a stirred solution 4-amino-5-chloro-2-methoxybenzoic acid (about 1.2parts by mole) and triethylamine (about 2.2 parts by mole) intetrahydrofuran (THF) was slowly added isobutyl chloroformate (about 1.2parts by mole) at room temperature. After about 30 minutes, a solutionof the piperidine ester (about 1.0 parts by mole) in THF was added tothe preformed, mixed anhydride. The reaction was stirred at roomtemperature for about 14 hours before diluting with a saturated solutionof sodium bicarbonate. The product was extracted out using, e.g., ethylacetate and the separated organic layer was further washed with brineand then dried over anhydrous sodium sulfate. Filtration andconcentration provided ATI-7505 free base.

ATI-7505 free-base was dissolved in ethanol and isopropanol (about 1:1ratio by volume) and cooled in an ice bath. To the ice cold solution wasslowly added concentrated hydrochloric acid and then warmed to roomtemperature. After about 7 hours of stirring at room temperature, thesolid was filtered and washed with ethanol and isopropanol (about 1:1ratio by volume) to provide a wet cake. The wet cake was resuspended inethanol and then heated to reflux. The stirred solution was warmed toroom temperature and allowed to recrystallize. The product was filteredunder vacuum rinsed with ethanol and then dried under vacuum to provideATI-7505 dihydrochloride salt was a white solid.

Method 4:

Alternate synthesis of (R)-quinuclidin-3-yl6-((3S,4R)-4-(4-amino-5-chloro-2-methoxybenzamido)-3-methoxypiperidin-1-yl)hexanoatedihydrochloride salt—ATI-7505 dihydrochloride salt:

To a mixture of (3R)-quinuclidinol in dichloromethane was addeddrop-wise 6-bromohexanoyl chloride. The reaction mixture was heated toreflux and after about 18 hours the reaction was cooled to roomtemperature. The unreacted (3R)-quinuclidinol was filtered away and tothe filtrate was added diethyl ether to precipitate out the desiredproduct. The product was filtered under vacuum and washed with CH₂Cl₂and diethyl ether (about 1:1 ratio by volume) to provide the product asa white solid.

Dibenzylamino piperidine (about 1.0 parts by mole) was added to amixture of 6-bromoalkanoyl ester (about 1.0 parts by mole) and potassiumcarbonate (about 2.2 parts by mole) in DMF solvent. The reaction mixturewas stirred at about 70° C. for approximately 11 hours before cooling toroom temperature and then diluted with a saturated solution of sodiumbircarbonate. The product was extracted out with ethyl acetate.Subsequent washing with brine, drying over anhydrous sodium sulfate,filtering and concentrating provided the product as colorless oil.

To a reaction flask containing palladium on carbon was added a solutionof the above dibenzyl piperidine ester (about 1.0 parts by mole) inmethanol, and to this mixture was added ammonium formate (about 4 partsby mole). The reaction was heated to reflux and after about 10 hours,the reaction flask was cooled to room temperature and the palladium oncarbon was filtered away, e.g., through a pad of celite. The filtratewas concentrated to an oil, which was purified via flash columnchromatography (e.g., SiO₂: about 150:10:1; CH₂Cl₂:CH₃OH:NH₄OH) toprovide the amino piperidine ester product as a yellow oil.

To a stirred solution 4-amino-5-chloro-2-methoxybenzoic acid (about 1.2parts by mole) and triethylamine (about 2.2 parts by mole) intetrahydrofuran (THF) was slowly added isobutyl chloroformate (about 1.2parts by mole) at room temperature. After about 30 minutes, a solutionof the piperidine ester (about 1.0 parts by mole) in THF was added tothe preformed, mixed anhydride. The reaction was stirred at roomtemperature for about 14 hours before diluting with a saturated solutionof sodium bicarbonate. The product was extracted out using, e.g., ethylacetate and the separated organic layer was further washed with brineand then dried over anhydrous sodium sulfate. Filtration andconcentration provided ATI-7505 free base.

ATI-7505 free-base was dissolved in ethanol and isopropanol (about 1:1ratio by volume) and cooled in an ice bath. To the ice cold solution wasslowly added concentrated hydrochloric acid and then warmed to roomtemperature. After about 7 hours of stirring at room temperature, thesolid was filtered and washed with ethanol and isopropanol (about 1:1ratio by volume) to provide a wet cake. The wet cake was resuspended inethanol and then heated to reflux. The stirred solution was warmed toroom temperature and allowed to recrystallize. The product was filteredunder vacuum rinsed with ethanol and then dried under vacuum to provideATI-7505 dihydrochloride salt was a white solid.

Method 5:

Alternate synthesis of (R)-quinuclidin-3-yl6-((3S,4R)-4-(4-amino-5-chloro-2-methoxybenzamido)-3-methoxypiperidin-1-yl)hexanoatedihydrochloride salt—ATI-7505 dihydrochloride salt:

Benzyl bromide (about 1.2 parts by mole) was added to a solution of(3R)-quinuclidinol (about 1.0 parts by mole) in dichloromethane. Thereaction was stirred at room temperature for about 4 hours beforefiltering and rinsing with dichloromethane to provide the product as awhite solid.

6-Bromohexanoyl chloride (about 1.1 parts by mole) was added to asolution of benzyl protected (3R)-quinuclidinol (about 1.0 parts bymole) and the reaction mixture was heat to about 60° C. After about 12hours, the reaction was cooled to room temperature and the product wasprecipitated out by the addition of diethyl ether. Following vacuumfiltration and rinsing with ether and drying provided the product as anamorphous solid.

A mixture of the piperidine (about 1.0 parts by mole), thealkanoylhalide ester (about 1.0 parts by mole) and triethylamine (about2.0 parts by mole) in DMF solvent was heated at about 60° C. for about 6hours. The reaction was then cooled to room temperature and diluted witha saturated solution of sodium bicarbonate and extracted over ethylacetate. Following brine wash and drying over anhydrous sodium sulfatethe organic layer was concentrated to provide the product as clear oil.

To a reaction flask containing palladium on carbon was added a solutionof the dibenzyl piperidine ester (about 1.0 parts by mole) in methanoland to this mixture was added ammonium formate (about 4 parts by mole).The reaction was heated to reflux and after about 10 hours. The reactionflask was then cooled to room temperature and the palladium on carbonwas filtered, e.g., through a pad of celite. The filtrate wasconcentrated to give an oil and was subsequently purified via flashcolumn chromatography (SiO₂: about 150:10:1; CH₂Cl₂:CH₃OH:NH₄OH) toprovide the amino piperidine ester product as a yellow oil.

To a stirred solution 4-amino-5-chloro-2-methoxybenzoic acid (about 1.2parts by mole) and triethylamine (about 2.2 parts by mole) intetrahydrofuran (THF) was slowly added isobutyl chloroformate (about 1.2parts by mole) at room temperature. After about 30 minutes, a solutionof the piperidine ester (about 1.0 parts by mole) in THF was added tothe preformed, mixed anhydride. The reaction was stirred at roomtemperature for about 14 hours before diluting with a saturated solutionof sodium bicarbonate. The product was extracted out using, e.g., ethylacetate and the separated organic layer was further washed with brineand then dried over anhydrous sodium sulfate. Filtration andconcentration provided ATI-7505 free base.

ATI-7505 free-base was dissolved in ethanol and isopropanol (about 1:1ratio by volume) and cooled in an ice bath. To the ice cold solution wasslowly added concentrated hydrochloric acid and then warmed to roomtemperature. After about 7 hours of stirring at room temperature, thesolid was filtered and washed with ethanol and isopropanol (about 1:1ratio by volume) to provide a wet cake. The wet cake was resuspended inethanol and then heated to reflux. The stirred solution was warmed toroom temperature and allowed to recrystallize. The product was filteredunder vacuum rinsed with ethanol and then dried under vacuum to provideATI-7505 dihydrochloride salt was a white solid.

Method 6

Alternate synthesis of (R)-quinuclidin-3-yl6-((3S,4R)-4-(4-amino-5-chloro-2-methoxybenzamido)-3-methoxypiperidin-1-yl)hexanoatedihydrochloride salt—ATI-7505 dihydrochloride salt. While specificreaction conditions are recited below for an exemplary synthesis underMethod 6, these specifics are not to be construed as limiting the scopeof the method. One of skill in the art will recognize that alterationsin reaction conditions, including but not limited to reaction times,temperatures and solvents used, may be made under the method. Reactionyields, where indicated, are also exemplary and therefore may vary foreach run and set of reaction conditions.

The synthesis of ATI-7505 from cis-APM tartrate was based on 9.7 g labrun procedure.

a. Synthesis C2

Raw Materials

-   -   cis-piperidine carbamate, 24 Kg    -   benzyl bromide, 37.8 Kg    -   KI, 1.67 Kg    -   K₂CO₃, 48.7 Kg    -   N-methylpyrrolidone (NMP), 200 Kg    -   EA (ethyl acetate), 360 Kg    -   water, 600 kg    -   isopropyl alcohol (IPA)/water (1:1 w/w), 250 Kg

Procedure

Charge cis-piperidine carbamate (24 Kg, 1 eq.) and K₂CO₃ (48.7 Kg, 6eq.) to a reactor, followed by adding NMP (200 Kg) to the reactor. Stirthe mixture at room temperature for 15 minutes. Add KI (1.67 Kg, 0.1eq.) to the reactor, followed by adding benzyl bromide (37.8 Kg, 2.2 eq)and increasing the temperature to 75° C. within 60 minutes. Sample thereaction mixture after about 4 hours; the expected reaction time isabout 7-9 hours.

After the reaction is deemed completed, add water (350 Kg) to thereactor and extract with EA (120 Kg; 3 times). Collect the EA layer andwash the EA layer with water (200 Kg; 3 times). Concentrate the EA layerto a solid at 70° C. Add IPA/water (1:1, 200 Kg) to the reactor and heatthe reactor to about 75-80° C. Add 25 Kg portions of IPA/water (1:1) at75-80° C. until a clear solution obtained. Slowly cool down the reactionmixture to 5° C. Collect the solid by filtration and dry the wet cake atabout 60° C. to obtain C2 (31.7 Kg; 82% yield). HPLC purity for theexemplary batch was 99.3%

b. Synthesis of C3

Raw Material

-   -   KOH, 56.3 Kg    -   IPA, 200 Kg    -   DCM (dichloromethane), 550 Kg    -   Water, 1300 Kg    -   C2, 32 Kg

Procedure

Add C2 (32 Kg, 1 eq), KOH (56.3 Kg, 12 eq) and IPA (200 Kg) to thereactor. Heat the reaction mixture to reflux temperature (about 82° C.).Sample the reaction after about 4 hours; the expected reaction time isabout 4-5 hours. After the reaction is completed, remove the IPA bydistillation at 50° C. Add DCM (230 Kg) and water (700 Kg) to thereactor and collect both layers. Back extract the water layer with DCM(160 K; 2 times) and combine the DCM layers. Wash the DCM layer withwater (200 Kg; 3 times) and concentrate the DCM layer at 70° C. toobtain C3 as an oil and proceed to the next step without isolation(assume a 100% yield).

c. Synthesis of Cis-AMP tartrate salt

Raw Materials

Add methanol (260 Kg) and water (130 Kg) to the reactor that contains C3from the previous step. Add (+)-DBT (15.2 Kg), which has been dissolvedin 130 Kg of methanol to the reactor at 70° C., preferably within 60minutes. Add a portion of methanol (70 Kg) to make ensure a clearsolution is obtained before cooling down the reaction mixture to 50° C.The product will come out around 50° C.; slowly cool down to about 10°C. before filtration. Collect solid by filtration and, preferably, checkthe enantiomeric excess (ee) value and solid content.

Place the solid in a reactor. Add MeOH/water (5:1, 600 Kg) to thereactor and heat the mixture to 70° C. More MeOH/water (5:1) can beadded to obtain a clear solution before cooling down the reactionmixture to 50° C. Slowly cool down the mixture to 10° C. beforecollecting the solid by filtration. Dry the wet cake at about 60° C. Inthis exemplary run, cis-AMP ½ (+)-DBT (12.6 Kg, 31% weight yield and 62%theoretical yield) was obtained with a HPLC purity 99.8% and an 97.9%ee.

d. Synthesis of the cis-AMP free base

Raw Material

-   -   cis AMP ½ DBT, 10 g, 0.0279 mole    -   isopropyl ether (IPE), 50 ml    -   water, 50 ml    -   45% NaOH, 7.2 g, 0.18 mole

Procedure

The piperidine (+)-dibenzoyltartrate salt (10.00 g; 0.0279 mole) wassuspended in 30 ml water and 50 ml IPE with aggressive stirring. 45%sodium hydroxide (7.2 g; 0.18 mole) was added dropwise until the solidwas dissolved. The IPE layers were washed with water (10 ml; 2 times),and concentration provided a white solid compound of crude free base(7.20 g).

e. Synthesis of C5

Raw Material

-   -   free base, 7.2 g, 0.0232 mole    -   ethyl 6-bromo-hexanoate, 4.76 g, 0.0214 mole    -   potassium carbonate, 5.77 g, 0.0418 mole    -   potassium iodide, 1.39 g, 8.37 mmole    -   DMF (dimethyl formamide), 30 ml    -   isopropyl ether (IPE), 50 ml    -   water, 50 ml.

Procedure

Free base (7.2 g), ethyl 6-bromo-hexanoate (4.75 g; 0.0214 mole),potassium carbonate (5.77 g; 0.0418 mole), potassium iodide (1.39 g;8.37 mmole), and DMF (30 ml) were charged to a reactor. The reactionmixture was heated to 70° C. for 1 hour and was monitored by HPLC forthe completion of the reaction. After 1 hour, the reaction was cooled toroom temperature and quenched with 30 ml water and 50 ml IPE. The IPElayers were washed with water (10 ml; 2 times). Concentration provided ayellow oil of crude compound C5 (9.10 g).

f. Synthesis of C6

Raw Material

-   -   C5, 9.10 g, 0.0201 mole    -   (R)-3-quinuclidinol, 5.20 g, 0.0409 mole    -   Ti(OiP)₄ (titanium (IV) isopropoxide), 1.16 g, 4.08 mmole    -   toluene, 120 ml    -   isopropyl ether, 60 ml    -   water, 80 ml

Procedure

C5 (9.10 g; 0.0201 mole), (R)-3-quinuclidinol (5.20 g; 0.0409 mole),Ti(OiP)₄ (1.16 g; 4.08 mmole), and toluene (120 ml) were charged inreactor and with stirring. The reaction was equipped with a packingcolumn (24/40; 15 cm in length) and a short path (24/40), which washeated to distill out EtOH, IPA and toluene (oil bath temperature 160°C.). The reaction mixture was monitored by HPLC for the completion ofthe reaction. In this exemplary synthesis, the HPLC showed the startingmaterial had been completely consumed. Pressure was reduced tofacilitate removal of the toluene. The reaction was cooled to roomtemperature and quenched with 40 ml water and 60 ml IPE, followed bywashing the IPE layers with water (20 ml; 2 times) and concentrating,providing a yellow oil compound crude C6 (11.70 g).

g. Synthesis of C7

Raw Material

-   -   C6, 11.70 g, 0.0220 mole    -   5% Pd/C, 1.0 g    -   IPA, 30 ml

Procedure

C7 (11.70 g), 5% Pd/C (1.0 g) and IPA (30 ml) were charged inhydrogenation reactor (N₂ inert; H₂ at 5 atmospheres). The mixture wasstirred and heated in a 70° C. water bath for 7 hours. The reactionmixture was monitored by HPLC and TLC for the completion of thereaction, which showed that the starting material had been completelyconsumed. The reaction was cooled to room temperature and filteringthrough a pad of celite with IPA rinsing. The filtrate was concentratedto provide a 6.66 g crude oil of C7.

h. Synthesis of ATI-7505 base

Raw Material

-   -   4-amino-5-chloro-2-methoxy-benzoic acid, 5.0 g, 0.0249 mole    -   THF, 30 g    -   triethylamine, 4.7 g, 0.0465 mole    -   pivaloyl chloride, 2.7 g, 0.0225 mole    -   C7, 6.66 g, 0.0189 mole    -   diethyl ketone (DEK), 100 ml    -   32% HCL    -   water    -   45% NaOH

Procedure

Pivaloyl chloride (2.7 g; 0.0225 mol) was added dropwise to a solutionof the 4-amino-5-chloro-2-methoxy-benzoic acid (5.0 g; 0.0249 mol) andtriethylamine (4.7 g; 0.0465 mmol) in THF (20 g) at room temperature.The reaction turned cloudy upon addition and after 60 minutes to thispreformed mixed anhydride was added a solution of C7 (6.66 g; 0.0189mol) in THF (10 g) and allowed to stir at room temperature. HPLC and TLCshowed the starting material had been completely consumed.

The reaction was quenched with water (40 ml) and DEK (40 ml), and 32%HCl was added to pH=4.0. The combined organic layers were washed withwater (10 ml; 2 times) and the aqueous layer was collected. DEK (60 ml)was added to the aqueous layer and 45% NaOH was added to pH=12. Extract,separate and drain off the aqueous layer. The combined organic layerswere washed with water (10 ml; 2 times) and concentrated to provide12.01 g yellow oil ATI-7505 base.

i. Synthesis of ATI-7505

Raw Material

-   -   ATI-7505 base, 12.01 g    -   Ethanol (EtOH), 50 ml    -   IPA, 70 ml    -   32% HCL

Procedure

The 12.01 g crude product of ATI-7505 base was dissolved in 50 ml EtOH.Concentrated 32% HCl was added slowly with stirring to pH=4.1. Afterstirring about 16 hours, 50 ml IPA was added and the reaction wasstirred for 2 hours. The solid was filtered and rinsed with 20 ml IPA.The solid was dried to constant weight to provide 9.71 g of ATI-7505 asa white solid. HPLC purity was 98.65%.

EXAMPLE 1

Preparation of6-[4R-(4-amino-5-chloro-2-methoxy-benzoylamino)-3S-methoxy-piperidin-1-yl]-hexanoicacid 1-aza-bicyclo[2.2.2]oct-3′R-yl ester, dihydrochloride salt

Step 1: Resolution of Racemic Norcisapride

(−)-2,3-Dibenzoyl-L-tartaric acid ((−)-DBT, about 1 part by weight) wasdissolved in ethanol and filtered to remove residual particulates.Separately, racemic norcisapride (about 0.8 part by weight) wasdissolved in a mixture of ethanol and water and then filtered. Thefiltrate was heated to about 75° C. before adding the (−)-DBT solution.After stirring at this temperature for about 30 minutes, the mixture wasslowly cooled for several hours to about 5° C. and the product salt wascollected under vacuum filtration and washed with EtOH/H₂O mixture. Thewetcake was recrystallized from EtOH/H₂O by heating to about 79° C. andslow cooling to about 5° C. as before. The product was collected on avacuum filter and washed with EtOH/H₂O to give a wetcake.

The wetcake was suspended in water and the pH was adjusted to about 12using 7% (W/W) aq. NaOH. The resulting suspension was stirred for about3 hours at room temperature before filtering under vacuum and washingthe solid material with water and drying under vacuum. The product wasthen retreated with (−)-DBT to form the salt by the same generalprocedure described above. The isolated salt was then neutralized withaq. NaOH as described above. The product was isolated on a filter anddried as before to provide (+)-norcisapride base (about 0.25 parts byweight). The e.e. by chiral HPLC analysis was about 100%(+)-norcisapride. The optical rotation was about +5° (methanol; 25° C.and 589 nm), confirming the positive isomer of norcisapride.

Step 2: Coupling with Ethyl 6-bromohexanoate

(+)-Norcisapride (about 1 part by weight), potassium carbonate (about0.48 part by weight) and potassium iodide (about 0.063 part by weight)were suspended in anhydrous USP ethanol. Ethyl 6-bromohexanoate (about0.76 part by weight) was added slowly to the suspension at roomtemperature. The mixture was heated to reflux until completion of thereaction. Subsequent cooling to room temperature the reaction mixturewas filtered to remove, e.g., inorganic solids, and the filtrate wasconcentrated under reduced pressure to about one-half the volume. Theproduct was precipitated by slowly adding the crude material to coldwater (about 13 parts by weight) with rapid stirring. The precipitatewas filtered under vacuum and washed with water and then reprecipitatedtwice more by dissolution in anhydrous ethanol and slow addition intocold water as before. The resulting wetcake was washed with n-heptaneand resuspended in ethyl acetate and n-heptane (1:9; v/v) and stirredfor about 1 hour and before filtering and drying under vacuum to yield0.73 parts by weight of the coupled product as a white solid.

Step 3: Coupling with (R)-3-Quinuclidinol and Dihydrochloride SaltFormation

The ester (1 part by weight) and (R)-3-Quinuclidinol (about 1.12 part byweight) were suspended in toluene before slowly adding titanium (IV)ethoxide (about 0.5 part by weight) to the stirred suspension. Themixture was heated to about 91° C. under a stream of nitrogen, andpartial vacuum was applied to the flask through a distillation apparatusin order to azeotropically remove the ethanol. Additional toluene wasadded as needed to maintain a minimum solvent volume in the flask. Thereaction was considered complete after about 33 hours.

The mixture was cooled to about room temperature and extracted fivetimes with water. The organic layer was concentrated under reducedpressure and the resulting residue was redissolved in EtOH/iPrOH (about1:1 v/v) and then filtered through a 0.45 micron membrane filter toremove any particulates. Concentrated hydrochloric acid was added slowlyto the stirred filtrate to precipitate out the desired product as thedihydrochloride salt. The resulting suspension was stirred for severalhours at room temperature and collected under vacuum filtration andrinsed with EtOH/iPrOH (1:1; v/v) to provide 0.53 part by weight of thecrude product salt.

Crudedihydrochloride salt was resuspended in ethanol and heated toreflux before cooling to room temperature over about 1 hour. The productwas collected under vacuum filtration and rinsed with ethanol and thenair-dried. The solids were resuspended in ethanol and warmed to about55° C. to give a clear solution before adding warm isopropanol and theproduct was allowed to precipitate by slow cooling to room temperature.The resulting suspension was stirred for several hours before vacuumfiltering and rinsing with, e.g., isopropanol. The product was vacuumdried, initially at room temperature for several hours and then at about55° C. until a constant weight was achieved.

EXAMPLE 2

(+) and (−)-norcisapride can be made from its racemic mixture byresolution of the enantiomers using conventional means such as opticallyresolving acids, according to the method described in U.S. Pat. No.6,147,093, or in “Enantiomers, Racemates and Resolutions”, by J.Jacques, A. Collet, and S. H. Wilen (Wiley-Interscience, New York,N.Y.), or in S. H. Wilen et al., Tetrahedron (1977) 33:2725.

The 4 isomers can be obtained in low-mg amounts by using preparativecolumn chromatography followed by evaporation of the solvent. Thismethod is useful for preparing small amounts for analytical andcharacterization purposes. This is a standard separation method usedroutinely in analytical labs in order to isolate and characterizemetabolites.

Possible synthetic routes to Compound IV, Compound VI and (+)-CompoundII are described below using (+)-norcisapride as a starting material.The routes to Compound III, Compound V and (−)-Compound II are identicalexcept that they use (−)-norcisapride as a starting material.Preferably, however, the methods and processes of Methods 2-5 are usedto generate Compounds II-VI and the other compounds of the invention.More preferably, the methods of Methods 3-5 are used to make thecompounds of the invention.

EXAMPLE 3

Production of (+)-Compound II, ethyl ester

A equimolar mixture of (+)-norcisapride and ethyl 6-bromohexanoate (1equivalent each), a catalytic amount of KI, and K₂CO₃ (2 equivalents) inDMF is heated at about 60° C. for several hours or until TLC analysisindicates that the reaction is over. After cooling to room temperature,water is added and the mixture is extracted with EtOAc. The combinedorganic extracts are washed successively with water, 10% LiCl(aq)solution and brine, then dried over Na₂SO₄. Concentration gives(+)-compound II, ethyl ester.

Production of (+)-Compound II

A mixture of crude (+)-compound II, ethyl ester, from above (1 eq.), KOH(2M, 5 eq.) in MeOH and THF (enough to dissolve) is stirred at roomtemperature for approximately 1 to 2 hours. The MeOH and THF are removedunder vacuum, and the residue is diluted with water. Wash with anorganic solvent such as EtOAc. The aqueous layer is acidified to pH ˜5using HCl. The precipitate is filtered off and dried to give(+)-Compound II.

Production of Compound IV and Compound VI

A mixture of (+)-Compound 11 (1 eq.), (R)-(−)-3-quinuclidinol HCl salt(1 eq.), EDAC (1 eq.) and DMAP (1 eq.) in DMF is heated at around 50Covernight. After cooling and diluting with water, the mixture ispurified by chromatography or by crystallization to provide Compound IV.Similarly, using (S)-(+)-quinuclidinol, Compound VI is obtained.

The following compounds are prepared essentially according to methodsand procedures described above, particularly those of Methods 2-5 andmore preferably according to those of Methods 3-5. The compound nameswere generated using either ChemDraw Ultra version 8.03, which isavailable from Cambridgesoft Corporation or ACD Namepro software,version 6.0. Table of Compounds (3S)-1-azabicyclo[2.2.2]oct-3-yl6-{(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}hexanoate;(3S)-1-azabicyclo[2.2.2]oct-3-yl 6-{(3R,4S)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}hexanoate;(3R)-1-azabicyclo[2.2.2]oct-3-yl 6-{(3R,4S)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}hexanoate;8-methyl-8-azabicyclo[3.2.1]oct-3-yl 6-{(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}hexanoate;4-[({(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)amino]benzoic acid; methyl4-[({(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)amino]benzoate; methyl4-[({(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)amino]benzoate; methyl4-[({(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)amino]benzoate; ethyl4-[({(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)amino]benzoate; isopropyl4-[({(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)amino]benzoate; 2-methoxyethyl4-[({(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)amino]benzoate; 2-pyrrolidin-1-ylethyl4-[({(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)amino]benzoate; 1-methylpiperidin-4-yl4-[({(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)amino]benzoate; 2-pyridin-2-ylethyl4-[({(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)amino]benzoate; 2-(dimethylamino)ethyl4-[({(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)amino]benzoate; 1-methylpiperidin-3-yl4-[({(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)amino]benzoate; 2-morpholin-4-ylethyl4-[({(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)amino]benzoate; 1,4-dimethylpiperidin-4-yl4-[({(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)amino]benzoate;4-[({(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)amino]benzoic acid; 2-oxo-2-(piperidin-4-ylamino)ethyl4-[({(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)amino]benzoate;1-({(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)piperidine-4-carboxylic acid; methyl1-({(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)piperidine-4-carboxylate; methyl1-({(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)piperidine-4-carboxylate; methyl1-({(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)piperidine-4-carboxylate; ethyl1-({(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)piperidine-4-carboxylate; 2-methoxyethyl1-({(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)piperidine-4-carboxylate;4-{[(2-{(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}ethyl)(methyl)amino]methyl}benzoic acid; methyl4-{[(2-{(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}ethyl)(methyl)amino]methyl}benzoate; methyl4-{[(2-{(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}ethyl)amino]methyl}benzoate; isopropyl4-{[(2-{(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}ethyl)amino]methyl}benzoate; ethyl4-{[(2-{(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}ethyl)amino]methyl}benzoate Dihydrochloride;(3R)-1-azabicyclo[2.2.2]oct-3-yl 4-{[(2-{(3S,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}ethyl)amino]carbonyl}benzoate;(R)-quinuclidin-3-yl6-((3S,4R)-4-(4-amino-5-chloro-2-methoxybenzamido)-3-methoxypiperidin-1-yl)hexanoate; or6-((3S,4R)-4-(4-amino-5-chloro-2-methoxybenzamido)-3-methoxypiperidin-1-yl)hexanoicacid

Formulation, Administration, and Uses

Dosage rates and routes of administration of the disclosed compounds aresimilar to those already used in the art and known to the skilledartisan (see, for example, Physicians' Desk Reference, 54th Ed., MedicalEconomics Company, Montvale, N.J., 2000).

The magnitude of a prophylactic or therapeutic dose of structural and/orfunctional analog of cisapride in the acute or chronic management ofdiseases and/or disorders described herein will vary with the severityof the condition to be treated, and the route of administration. Thedose, and perhaps the dose frequency, will also vary according to theage, body weight, and response of the individual patient. In general,the total daily dose range for structural and/or functional analogs ofcisapride, for the conditions described herein, is from about 1 mg toabout 200 mg, in single or divided doses. Preferably, a daily dose rangeshould be between about 5 mg to about 100 mg, in single or divideddoses, while most preferably, a daily dose range should be between about5 mg to about 75 mg, in single or divided doses. It is preferred thatthe doses are administered from 1 to 4 times a day. In managing thepatient, the therapy should be initiated at a lower dose, perhaps about5 mg to about 10 mg, and increased up to about 50 mg or higher dependingon the patient's global response. It is further recommended thatchildren, and patients over 65 years, and those with impaired renal orhepatic function, initially receive low doses, and that they be titratedbased on individual response(s) and blood level(s). It may be necessaryto use dosages outside these ranges in some cases as will be apparent tothose skilled in the art. Further, it is noted that the clinician ortreating physician will know how and when to interrupt, adjust, orterminate therapy in conjunction with individual patient response.

The compounds of the subject invention can be formulated according toknown methods for preparing pharmaceutically useful compositions.Formulations are described in detail in a number of sources which arewell known and readily available to those skilled in the art. Forexample, Remington's Pharmaceutical Science by E. W. Martin describesformulations which can be used in connection with the subject invention.In general, the compositions of the subject invention are formulatedsuch that an effective amount of the bioactive compound(s) is combinedwith a suitable carrier in order to facilitate effective administrationof the composition.

The compositions of the subject invention include compositions such assuspensions, solutions and elixirs; aerosols; or carriers such asstarches, sugars, microcrystalline cellulose, diluents, granulatingagents, lubricants, binders, disintegrating agents, and the like, in thecase of oral solid preparations (such as powders, capsules, and tablets)with the oral solid preparations being preferred over the oral liquidpreparations. A preferred oral solid preparation is capsules. The mostpreferred oral solid preparation is tablets. Preferred amounts of activeingredient (i.e., an structural and/or functional analog of cisapride)in a solid dosage form are about 5 mg, 10 mg, and 25 mg.

Further, acceptable carriers can be either solid or liquid. Solid formpreparations include powders, tablets, pills, capsules, cachets,suppositories and dispersible granules. A solid carrier can be one ormore substances which may act as diluents, flavoring agents,solubilizers, lubricants, suspending agents, binders, preservatives,tablet disintegrating agents or encapsulating materials.

The disclosed pharmaceutical compositions may be subdivided into unitdoses containing appropriate quantities of the active component. Theunit dosage form can be a packaged preparation, such as packetedtablets, capsules, and powders in paper or plastic containers or invials or ampules. Also, the unit dosage can be a liquid basedpreparation or formulated to be incorporated into solid food products,chewing gum, or lozenge.

In addition to the common dosage forms set out above, the compounds ofthe present invention may also be administered by controlled releasemeans and/or delivery devices such as those described in U.S. Pat. Nos.:3,845,770; 3,916,899; 3,536,809; 3,598,123; and 4,008,719, thedisclosures of which are hereby incorporated by reference in theirentirety.

Any suitable route of administration may be employed for providing thepatient with an effective dosage of a structural and/or functionalanalog of cisapride. For example, oral, rectal, parenteral(subcutaneous, intramuscular, intravenous), transdermal, and like formsof administration may be employed. Dosage forms include tablets,troches, dispersions, suspensions, solutions, capsules, patches, and thelike.

One aspect of the invention provides methods and/or processes for makingthe compounds and compositions of the invention.

One aspect of the invention provides a method of treatinggastroesophageal reflux disease in a mammal, while substantiallyreducing the concomitant adverse effects associated with theadministration of cisapride, which comprises administering to a human inneed of such treatment, a therapeutically effective amount of astructural and/or functional analog of cisapride, or a pharmaceuticallyacceptable salt thereof. A preferred aspect is the treatment ofgastroesophageal reflux disease in humans.

Another aspect of the invention provides a composition for the treatmentof a human suffering from gastroesophageal reflux disease, whichcomprises a therapeutically effective amount of a structural and/orfunctional analog of cisapride, or a pharmaceutically acceptable saltthereof.

Yet another aspect of the present invention provides a method ofeliciting an anti-emetic effect in a mammal, while substantiallyreducing the adverse effects associated with the administration ofcisapride, which comprises administering to a mammal in need of suchanti-emetic therapy, a therapeutically effective amount of structuraland/or functional analogs of cisapride, or a pharmaceutically acceptablesalt thereof. Preferably, the mammal is a human.

In an additional aspect, the present invention encompasses ananti-emetic composition for the treatment of a mammal in need ofanti-emetic therapy, which comprises a therapeutically effective amountof a structural and/or functional analog of cisapride, or apharmaceutically acceptable salt thereof.

A further aspect of the present invention includes a method of treatinga condition caused by gastrointestinal motility dysfunction in a mammalwhich comprises administering to a mammal in need of treatment forgastrointestinal motility dysfunction, a therapeutically effectiveamount of a structural and/or functional analog of cisapride, or apharmaceutically acceptable salt thereof. Conditions caused bygastrointestinal motility dysfunction include, but are not limited to,dyspepsia, gastroparesis, constipation, post-operative ileus, andintestinal pseudo-obstruction. Preferably, the mammal is a human.

The observation that cisapride enters the central nervous system andbinds to 5HT₄ receptors indicates that cisapride may havecentrally-mediated effects. Cisapride is a potent ligand at 5HT₄receptors, and these receptors are located in several areas of thecentral nervous system. Modulation of serotonergic systems has a varietyof behavioral effects. Accordingly, the compounds of the subjectinvention can be used in the treatment of: 1) cognitive disorders,including but not limited to Alzheimer's disease; 2) behavioraldisorders, including but not limited to schizophrenia, mania,obsessive-compulsive disorder, and psychoactive substance use disorders;3) mood disorders, including but not limited to depression and anxiety;and 4) disorders of control of autonomic function, including but notlimited to essential hypertension and sleep disorders.

Accordingly, the present invention also provides methods of treatingcognitive, behavioral, mood, or autonomic function control disorders ina mammal comprising the administration of a therapeutically effectiveamount of structural and/or functional analog of cisapride, or apharmaceutically acceptable salt thereof. Preferably, the mammal is ahuman.

ATI-7505 Binds with High Affinity to 5-HT₄ Receptors

The 5-HT₄ receptor is known to be the major receptor subtype involved inthe prokinetic activity of cisapride in the gut. ATI-7505 has a highbinding affinity for 5-HT₄ receptor, with a low nanomolar IC₅₀. As shownin Table 1, the affinity of ATI-7505 for the 5-HT₄ receptor was 18-foldgreater than cisapride and at least 360-fold greater than the majormetabolite of ATI-7505, the carboxylic acid. TABLE 1 5-HT₄ ReceptorBinding 5-HT₄ Receptor Guinea Pig Striatum Compound IC₅₀ (nM) K_(i) (nM)n_(H) ATI-7505 8.3 1.4 0.7 ATI-7500 >3,000 >500 — Cisapride 150 24.9 0.8n_(H), Hill coefficient.5-HT₄ receptor prototypic reference antagonist [³H]GR113808 (0.70 nM)

ATI-7505 is a Highly Potent Partial Agonist at Human 5-HT₄ Receptor

ARYx performed in vitro assays based on adenylyl cyclase stimulation incells engineered to stably express human 5-HT₄ receptor. ATI-7505 provedto be a highly potent 5-HT₄ receptor agonist, whereas its majormetabolite, ATI-7500 was relatively weak (FIG. 1 and Table 2). Theestimated EC₅₀ of ATI-7505 (4 nM) was approximately 10-fold lower thanthat of cisapride (49 nM), and approximately 100-fold lower than that ofATI-7500 (395 nM). Based on its estimated Emax value, ATI-7505 had 85%of the efficacy of 5-HT (serotonin) (Table 2), demonstrating thatATI-7505 is a partial agonist of HT₄ receptors. TABLE 2 Potency andEfficacy (Intrinsic Activity) at Human 5-HT₄ Receptor Potency EfficacyCompound EC₅₀ pEC₅₀ % of 5HT (serotonin) 5-HT (serotonin) 46 7 NAATI-7505 4 8.45 85 ATI-7500 395 6.40 81 Cisapride 49 7 77EC₅₀, concentration causing 50% maximal increase in adenylyl cyclaseactivitypEC₅₀, negative logarithm of the EC₅₀

ATI-7505 Accelerates Gastric Emptying in Fed Dogs To characterize theeffects of ATI-7505 on gastric emptying, experiments were performed in apost-prandial model involving conscious dogs instrumented with sets ofstrain gauge transducers placed on the stomach and small bowel. Theobjective of the experiments was to measure the time required formigrating motor contractions (MMCs) to return to baseline followingingestion of a solid meal. A drug-induced shortening of MMC return timeindicated an early end of the digestive period due to acceleratedgastric emptying. Immediately after completion of an MMC in themid-small intestine, various doses of test drugs (vehicle, ATI-7505, orcisapride) were infused intravenously (IV) over 20 minutes. At the endof the drug infusion, the dogs were fed a meal. Gut contractions wererecorded for a minimum of 60 minutes prior to the start of the druginfusion to establish the fasting state and to identify the onset of MMCin the duodenum, and at least 30 minutes after the return of MMC in theduodenum. Quantitative comparisons of the treatments were based on thetime of MMC return as an index of gastric emptying following ingestionof a solid meal. As summarized in FIG. 2, ATI-7505 significantlyshortened the time of MMC return, indicating an acceleration of gastricemptying in normal fed dogs. Cisapride showed a similar pattern ofaction.

ATI-7505 Increases Gastric and Small Intestinal Motor Activity withNegligible Effect on Colonic Activity

Experiments were performed in fasted, conscious dogs to evaluate thegastric, small intestinal and colonic motor activity of ATI-7505compared to cisapride. A specific goal was to determine the dose sizesof ATI-7505 (IV and PO) that most closely mimic the pattern andmagnitude of contractile activity caused by cisapride at typicaltherapeutic doses in dogs (0.5 mg/kg IV; 1 mg/kg PO).

When given IV and PO, both ATI-7505 and cisapride caused prokineticeffects in the dog gut. The onset of action typically occurred within 1to 2 minutes and 25 to 30 minutes following IV and PO administration,respectively. The effect of ATI-7505 on gastric and small intestinalmotor activity mimicked cisapride. Like cisapride, ATI-7505 appeared tocause dose-dependent stimulation of antral and small bowel contractilitywith relatively little effect on colonic motor activity. The prokineticeffects caused by ATI-7505 in the upper GI tract occurred along with asmall but significant (p<0.05) increase in the frequency of giantmigrating contractions (GMC).

ATI-7505 was not associated with the development of retrograde giantmigrating contractions (RGC). Like cisapride, ATI-7505 had a minimaleffect on migrating motor complex (MMC) characteristics in the antrum aswell as proximal, mid and distal small intestine. With regard to MMCfrequency and phase III duration, only one significant difference wasnoted: PO ATI-7505 increased MMC frequency in the proximal smallintestine relative to the controls. The dogs tolerated the IV and POdoses of ATI-7505 well and exhibited no side effects such as diarrhea,anorexia, or weight loss.

Overall, the results showed that on a mg/kg-basis, ATI-7505 wasapproximately twice as potent as cisapride. In addition, the actions ofATI-7505, like those of cisapride, were consistent with a mechanisminvolving the facilitation of acetylcholine release from enteric neuronsrather than a direct smooth muscle action. In conclusion, ATI-7505increases gastric and small intestinal motor activity in acisapride-like manner with minimal-to-no effect on colonic activity.

The Metabolism of ATI-7505 is CYP450-Independent

Based on data from pooled human microsomes, ATI-7505 undergoesbiotransformation to a single metabolite, ATI-7500, which does notappear to be subject to further metabolism. The conversion of ATI-7505to ATI-7500 was not dependent on the presence of NADPH. Thus the majorbiotransformation pathway for ATI-7505 occurs independently of CYP450enzymes.

ATI-7505 Does Not Inhibit CYP450 Enzymes

To test the potential for ATI-7505 and/or its main metabolite, ATI-7500to act as CYP450 inhibitors, these two molecules were screened usingGentest Supersomes™. Consistent with published reports, cisapride hadsignificant inhibitory activity against CYP450 enzyme isoforms, CYP3A4,2D6 and to a lesser extent 2C9. Neither ATI-7505 nor its primarymetabolite, ATI-7500 displayed significant inhibitory activity againstthese three CYP450 isoforms, nor against a panel of other isoforms knownto play a role in drug metabolism.

ATI-7505 Has Negligible Affinity for the Cardiac Channel, I_(Kr)

The rapidly activating delayed rectifier potassium (K+) current inhumans (human I_(Kr)) is a K+ channel that is encoded by thehuman-ether-a-go-go-related gene (hERG). Cisapride is known to produceQT interval prolongations via a blockade of I_(Kr), and it was thereforeof interest to determine if ATI-7505 and ATI-7500 have importantinhibitory effects on human I_(Kr). The test system was mammalianHEK-293 cells expressing the hERG K+channels, in which the potassiumcurrent was measured by whole cell patch-clamp technique. The ranking ofthe IC₅₀ values was: cisapride (9.5 nM)>ATI-7505 (24,521 nM)>ATI-7500(204,080 nM) (Table 3). Overall, the findings indicate that ATI-7505 hasa significantly lower pro-arrhythmic potential than cisapride andsuggest that both ATI-7505 and ATI-7500 have negligible affinity forhuman I_(Kr) channels. TABLE 3 Inhibition of I_(Kr) Activity Activity ofI_(Kr) in HEK Cells % I_(Kr) control Compound (10,000 nM) IC₅₀ ATI-750578.0 24521 ATI-7500 88.9 204080 Cisapride 0 9.5Data are normalized to % control tail I_(Kr) (current elicited withoutdrug or vehicle present)

ATI-7505 Does not Induce Important Electrophysiological Changes inGuinea Pig Hearts

The cardiac electrophysiological effects of ATI-7505 were examined inisolated, perfused guinea pig hearts. The study examined ATI-7505,ATI-7500 and cisapride, all of which were each tested at concentrationsup to 10,000 nM. The no observed effect level (NOEL) was defined as thehighest concentration of test compound not showing a response that wassignificantly different from baseline (p<0.05). The following 6 cardiacparameters were tested: (1) QT interval; (2) MAPD₉₀; (3) SA interval;(4) QRS interval; (5) AH interval; and (6) HV. While ATI-7505 was a veryweak modulator of cardiac electrophysiologic parameters, its metabolite,ATI-7500 entirely lacked electrophysiological activity (Table 4). TheNOEL for ATI-7500 was>10,000 nM for the entire set of 6 cardiovascularparameters. Since cisapride had a NOEL of 10 nM for the combined set of6 cardiac parameters tested, while ATI-7505 had a combined NOEL of 1,000nM, ATI-7505 appears to lack the potency of cisapride in modulatingcardiac electrophysiologic parameters. Overall, the findings demonstratethat ATI-7505 is significantly safer than cisapride with regard to thepotential to induce important cardiac electrophysiologic fluctuations.TABLE 4 Cardiac Electrophysiologic Parameters in Isolated PerfusedElectrophysiological No Observed Effect Level (NOEL) Parameter CisaprideATI-7505 ATI-7500 QT Interval 10 1,000 >10,000 MAPD₉₀ 10 1,000 >10,000SA Interval 100 >10,000 >10,000 QRS Interval 1,000 >10,000 >10,000 AHInterval 1,000 >10,000 >10,000 HV Interval 1,000 1,000 >10,000 CombinedParameters 10 1,000 >10,000All molecules were tested at baseline, 10, 100, 1,000, and 10,000 nM.Other than for values reported as >10,000 nM, a significant difference(p < 0.05) from baseline was observed when the molecule was tested at a10-fold higher

Metabolism in Human Microsomal Preparations

The metabolism of these compounds was studied in pooled human microsomesin the presence and absence of the Cytochrome P-450 cofactor NADPH andboth the disappearance of parent and the appearance of the correspondingacid metabolite, i.e., the corresponding compound-II isomer, monitoredwith time.

As shown in Table 5, Compounds III and IV were rapidly hydrolyzed byesterase to their respective metabolites (+) and (−)-Compound II. Themetabolism was not dependent on CYP450 since the rate of hydrolysis wasindependent on NADPH presence, which is a necessary cofactor for CYP450to function. In contrast, (±)-S Compounds V and VI appeared to be quitestable with time under the same conditions. In this experiment, theamount of substrate (compounds III, IV, V, and VI) remaining in thereaction after 5, 60, and 90 minutes were evaluated by a tandem HPLC-MSmethod. This remaining amount was correlated with the appearance of themetabolite compound II. The sum of remaining substrate and compound IIwas constant over time and equal to the amount of starting material attime zero, therefore indicating that hydrolysis was the only metabolicreaction taking place. TABLE 5 test compounds were incubated in pooledhuman microsomal preparation in the presence of NADPH cofactor. Theremaining amount of test compound and the appearance of the metabolitecompound II were monitored over 90 minutes. Test compound CompoundsCompound III and IV V and VI Remaining Remaining Test Test CompoundMetabolite Compound Metabolite Time (ng/mL) (ng/mL) Sum (ng/mL) (ng/mL)Sum  5 31.3 2 33.3 32.9 1.5 34.4 60 20.7 14.5 35.2 29.9 1.5 31.4 90 16.919.4 36.3 31.9 1.5 33.4

Metabolism in Fresh Human Blood.

Test compounds were dissolved in DMSO to make 12.5 mM stock solution anddiluted with water to a final concentration of 2.5 mM (DMSO/H2O=20/80).Fresh blood was collected into heparinized tubes from 3 human donors andblood was stored on ice until incubation. Separate aliquots of bloodfrom each donor were pipetted into 1.5 mL centrifuge tubes and the tubeswere pre-incubated in a shaking water bath at 37° C. for 5 minutes. Thereaction was initiated by the addition of 10 μL of the appropriate testcompound stock to each tube (final concentration=100 μM). Incubationswere quenched after 0, 5, 15, 30 and 60 minutes, by the addition ofacetonitrile (750 mL), centrifuged at 12,000 rpm for 2 minutes and thesupernatant analyzed on an Agilent 1100 HPLC system. Separations wereaccomplished on a Keystone Intersil ODS2, 250×4.6 mm, 5 m column. Theaqueous mobile phase consisted of 20 mM ammonium acetate buffer (pH 5.7)and the organic phase acetonitrile. A gradient was used: initialcondition consisted of 20% acetonitrile for 1 minute. The acetonitrileconcentration was increased linearly to 90% over the next 8 minutes andheld there for 1 minute. The system was then recycled to initialconditions over the course of 1 minute and held there for 4 minutesbefore the next injection. The peak area for the parent peak wasdetermined by monitoring absorbance at 240, 254 and 290 nM. The resultswere expressed as amount of initial compound remaining and datasubjected to kinetic analysis using WinNonLin. The half-lives for theindividual compounds are given below in Table 6. TABLE 6 DiastereomericConfiguration Compound Norcis “half” Quinuclindol “half” Half-life (min)III − R Subject 1 12.03 Subject 2 10.37 Subject 3 9.23 Mean ± SD 10.5 ±1.41 IV + R Subject 1 8.47 Subject 2 8.61 Subject 3 8.58 Mean ± SD 8.59± 0.077 V − S Subject 1 >60 min Subject 2 >60 min Subject 3 >60 min VI +S Subject 1 >60 min Subject 2 >60 min Subject 3 >60 min

It should be understood that the examples and aspects described hereinare for illustrative purposes only and that various modifications orchanges in light thereof will be suggested to persons skilled in the artand are to be included within the spirit and purview of this applicationand the scope of the appended claims. Further, all patents, patentapplications, provisional applications, and publications referred to orcited herein are incorporated by reference in their entirety to theextent they are not inconsistent with the explicit teachings of thisspecification.

The invention and the manner and process of making and using it, are nowdescribed in such full, clear, concise and exact terms as to enable anyperson skilled in the art to which it pertains, to make and use thesame. It is to be understood that the foregoing describes preferredaspects of the invention and that modifications may be made thereinwithout departing from the spirit or scope of the invention as set forthin the claims. To particularly point out and distinctly claim thesubject matter regarded as invention, the following claims conclude thisspecification.

1. A method for preparing(R)-quinuclidin-3-yl6-((3S,4R)-4-(4-amino-5-chloro-2-methoxybenzamido)-3-methoxypiperidin-1-yl)hexanoateor a salt thereof, comprising: 1) optionally converting a compound offormula (I′)

 to a salt, wherein R is (C₁-C₈)alkyl; 2) converting the compound offormula (I′) or its salt to a compound of formula (II′)

 or its salt, respectively, wherein X₁ is a nitrogen protecting groupand X₂ is selected from the group consisting of hydrogen and a nitrogenprotecting group (wherein commonly known and used N protecting groupscan be used, e.g., N-benzyl; N-nitrobenzyl; N-BoC; N-oxide;N-paramethoxybenzyl; N-benzylsulfonyl); 3) treating the compound offormula (II′) with an alkali metal hydroxide or hydride to yield acompound of formula (III′)

4) producing a chiral salt of III′ by treating the compound of formula(III′) with a chiral resolving agent and isolating the chiral salt ofIII′; 5) optionally recrystallizing the product of 4; 6) basifying theproduct of 4 or 5 to yield the free base form of the product of 4 or 5;7) contacting the product of 6 with a (C₁-C₈)alkyl 6-halohexanoate toyield a compound of formula (IV′)

 wherein R′ is (C₁-C₈)alkyl; 8) treating the product of 7 with(R)-quinuclidin-3-ol and a Lewis acid in an organic solvent to yield acompound of formula (V′)

9) deprotecting the 4-amino group of the product of 8 to yield(R)-quinuclidin-3-yl6-[(3S,4R)-4-amino-3-methoxypiperidin-1-yl]hexanoate; 10) acylating theproduct of 9 with 4-amino-5-chloro-2-methoxybenzoic acid; 11) optionallyconverting the product of 10 to a salt.
 2. The process according toclaim 1 wherein R is (C₁-C₆)alkyl.
 3. The process according to claim 1wherein R is (C₁-C₄)alkyl.
 4. The process according to claim 1 wherein Ris ethyl.
 5. The process according to claim 4 wherein X₁ is benzyl andX₂ is benzyl.
 6. The process according to claim 5 wherein R′ is ethyl.7. The process according to claim 1 wherein the chiral salt is(+)-2,3-dibenzoyl-D-tartaric acid.
 8. The process according to claim 1wherein the deprotection is with H₂/Pd/C or ammonium formate/Pd/C. 9.The process according to claim 1 wherein the Lewis acid is a titaniumtetraalkoxide.
 10. The process according to claim 9 wherein the titaniumtetraalkoxide is Ti(OiP)4 (titanium (IV) isopropoxide).
 11. A processfor preparing (R)-quinuclidin-3-yl6-((3S,4R)-4-(4-amino-5-chloro-2-methoxybenzamido)-3-methoxypiperidin-1-yl)hexanoateor a salt thereof, comprising: 1) converting a compound that is ethyl4-amino-3-methoxypiperidine-1-carboxylate

 to a salt; 2) converting the ethyl4-amino-3-methoxypiperidine-1-carboxylate salt to ethyl4-(diphenylamino)-3-methoxypiperidine-1-carboxylate

3) treating the ethyl4-(diphenylamino)-3-methoxypiperidine-1-carboxylate with an alkali metalhydroxide or hydride to yield 3-methoxy-N,N-diphenylpiperidin-4-amine

4) preparing a chiral salt of the cis isomer of-methoxy-N,N-diphenylpiperidin-4-amine by contacting the3-methoxy-N,N-diphenylpiperidin-4-amine with a chiral resolving agentand isolating the chiral salt of the cis isomer of-methoxy-N,N-diphenylpiperidin-4-amine thereby produced; 5) optionallyrecrystallizing the product of 4; 6) basifying the product of 4 or 5 toyield the free base form of the product of 4 or 5

7) contacting the product of 6 with ethyl 6-bromohexanoate to yieldethyl 6-((3S,4R)-4-(diphenylamino)-3-methoxypiperidin-1-yl)hexanoate

8) esterifying the ethyl6-((3S,4R)-4-(diphenylamino)-3-methoxypiperidin-1-yl)hexanoate with(R)-quinuclidin-3-ol and a Lewis acid to yield (R)-quinuclidin-3-yl6-((3S,4R)-4-(diphenylamino)-3-methoxypiperidin-1-yl)hexanoate

9) deprotecting the 4-amino group of the product of 8 to yield(R)-quinuclidin-3-yl6-[(3S,4R)-4-amino-3-methoxypiperidin-1-yl]hexanoate; 10) acylating theproduct of 9 with 4-amino-5-chloro-2-methoxybenzoic acid to yield(R)-quinuclidin-3-yl6-((3S,4R)-4-(4-amino-5-chloro-2-methoxybenzamido)-3-methoxypiperidin-1-yl)hexanoate;11) optionally converting the product of 10 to a salt.
 12. The processaccording to claim 11 wherein the salt of step 1 is HCl.
 13. The processaccording to claim 11 wherein the alkali metal hydroxide of step 3 isKOH.
 14. The process according to claim 11 wherein the chiral salt ofstep 4 is (+)-2,3-dibenzoyl-D-tartaric acid.
 15. The process accordingto claims 11 wherein the Lewis acid is Ti(OiP)4 (titanium (IV)isopropoxide).
 16. The process according to claim 11 wherein thedeprotecting of the 4-amino group is conducted with H₂/Pd/C.
 17. Theprocess according to claim 11 wherein the acylation of the product of 9is conducted with pivaloyl chloride.
 18. A method for preparing acompound for formula II′

comprising converting a compound of formula (I′)

or its salt to a compound of formula (II′) or its salt, respectively,wherein X₁ is a nitrogen protecting group and X₂ is selected from thegroup consisting of hydrogen and a nitrogen protecting group, a R is(C₁-C₈)alkly.
 19. A compound of formula II′

or a salt thereof, wherein X₁ is a nitrogen protecting group and X₂ isselected from the group consisting of hydrogen and a nitrogen protectinggroup, and R is (C₁-C₈)alkyl.
 20. A method of making a compound offormula III′

comprising a treating compound of formula (II′)

with an alkali metal hydroxide or hydride, wherein X₁ is a nitrogenprotecting group and X₂ is selected from the group consisting ofhydrogen and a nitrogen protecting group, and R is (C₁-C₈)alkyl.
 21. Acompound of formula III′

or a salt thereof, wherein X₁ is a nitrogen protecting group and X₂ isselected from the group consisting of hydrogen and a nitrogen protectinggroup.
 22. A method of making a compound of formula III″

comprising a) contacting a compound of formula III′

 with a chiral resolving agent to yield a chiral salt of III″ andseparating the chiral salt of III″; b) optionally recrystallizing theproduct of a); and c) contacting the product of a) or b) with a base toyield the compound for formula III″; wherein X₁ is a nitrogen protectinggroup and X₂ is selected from the group consisting of hydrogen and anitrogen protecting group.
 23. The method of claim 22 wherein the chiralresolving agent is (+)-2,3-dibenzoyl-D-tartaric acid and the salt ofIII′ is a (3S,4R)-enantiomer (+)-2,3-dibenzoyl-D-tartrate salt.
 24. Themethod of claim 23 wherein one equivalent of(+)-2,3-dibenzoyl-D-tartaric acid is used for two equivalents of III′.25. A compound of formula III″

or a salt thereof, wherein X₁ is a nitrogen protecting group and X₂ isselected from the group consisting of hydrogen and a nitrogen protectinggroup.
 26. A method of making a compound of formula IV′

comprising contacting a compound for formula

with a (C₁-C₈)alkyl 6-halohexanoate, wherein R′ is (C₁-C₈)alkyl(preferably ethyl), and X₁ is a nitrogen protecting group and X₂ isselected from the group consisting of hydrogen and a nitrogen protectinggroup.
 27. A compound of formula IV′

or a salt thereof, wherein R′ is (C₁-C₈)alkyl (preferably ethyl), and X₁is a nitrogen protecting group and X₂ is selected from the groupconsisting of hydrogen and a nitrogen protecting group.
 28. A method ofmaking a compound of formula V′

comprising contacting a compound of formula IV′

with (R)-quinuclidin-3-ol and a Lewis acid in an organic solvent,wherein R′ is (C₁-C₈)alkyl and X₁ is a nitrogen protecting group and X₂is selected from the group consisting of hydrogen and a nitrogenprotecting group.
 29. The method according to claim 28 wherein the Lewisacid is a titanium tetraalkoxide.
 30. The method according to claim 29wherein the titanium tetraalkoxide is Ti(OiPr)₄ (titaniumtetraisopropoxide).
 31. A compound of formula V′

or a salt thereof, wherein X₁ is a nitrogen protecting group and X₂ isselected from the group consisting of hydrogen and a nitrogen protectinggroup.
 32. A method of making a compound of formula VI′

comprising cleaving groups X₁ and X₂ from a compound of formula V′

wherein X₁ is a nitrogen protecting group and X₂ is selected from thegroup consisting of hydrogen and a nitrogen protecting group.
 33. Themethod according to claim 32, wherein X₁ and X₂ are benzyl and arecleaved by treatment of V′ with H₂/Pd/C.
 34. A method of making(R)-quinuclidin-3-yl6-((3S,4R)-4-(4-amino-5-chloro-2-methoxybenzamido)-3-methoxypiperidin-1-yl)hexanoate(VII′) comprising contacting a compound of formula VI′

with 4-amino-5-chloro-2-methoxybenzoic acid
 35. The method of claim 34wherein said contacting is done in the presence of a pivaloyl halide.36. The method of claim 35 wherein the pivaloyl halide is pivaloylchloride.